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OfflineMephistophelian
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Mephisto's Master Slant Tek * 24
    #12968316 - 07/28/10 06:26 PM (13 years, 9 months ago)

Its easy...doesn't take special equipment or nonsense and is very straightforward. It also works with any type of slant you may take interest in be it PP Screw Cap, Simport CulTubes, Pyrex Screw Cap, or Vac-Tainers. This works with them all...especially if you don't have a sterilizer setup or a test-tube rack.

Tube Measurements:

  • Vac-Tainers - with a red top are commonly 10ml and 16mm x 100mm
  • CulTubes - depends on your model, I used the largest being 17mm x 100mm (They are also PP)
  • Absolute Smallest Recommended Dimensions is 16mm x 100mm, no less.


So lets begin.

Materials Needed:
  • Wood Sticks - Popsicle, Match Stick, whatever type you find that is flat and suitable for the job
  • Slants/Tubes w/ Caps- Vac-Tainer, CulTubes, Pyrex or PP
  • Agar Mix - I used Lab Standard MEA
  • Measuring Container - Mix agar
  • Syringe - Whatever size & gauge you have laying around.
  • Pint Jars - Quart jars would work as well, although I wouldn't want to personally do it with Quart jars
  • Cooking Pot - Any size is suffice Optional
  • Scale - For measuring agar mix (unless you have premix)






Step 1: Preparation

First thing, measure your sticks to see if they fit sufficiently. I had to break all my sticks in half lengthwise then in half width wise to fit right. Do this before anything else. Pre-soak your woodsticks. I boiled some water in a pot and dropped them in. Took it off the stove, covered with a lid and let soak. You can move onto step two while you wait.




Step 2: Preparation Continued

Fill your slant with water WITH a stick in it, to approximately 7ml. No more then that. As low as 5ml but no more then 7ml WITH the stick in it. Without the stick, it will vary by .5ml. You need this to be no more then 7ml because when you go to lay your slant down, you will lose surface area for your mycelium to grow. That's no good. You'll see when you pour your water in and slant it so the water doesn't go past the edge (remember that the lid will take up a small space inside the tube and you don't want agar building a wall up against it inside).



Now, you want to go into your Lab/Room and inside your GB or in front of your flow hood, have your spot where your slants will sit while they cool before hand. It sucks to do it while your unloading the jars and getting it ready last minute. Not wise. We see here that my little racks sitting on 4 popsicle sticks was suffice to make the perfect slant.



Now that we have that setup, lets go check on our sticks sitting in the hot water.

Step 3: Sticks 'An Tubes



Looks like the sticks are ready and saturated. In a pinch, you can soak these overnight instead of hot soaking, but hey...sometimes you forget or get an urge to do something right-this-second and well hot soak works well. Now what you do is get your tubes ready and lay them out and put your sticks in them. Once you've got your sticks laid out, and sticks in them. We can go to the next step. See how easy this is? You could be drinking a beer while doing this. But make sure your still on your first by this point otherwise your in for a world of trouble later. :wink:


Step 4: Agar Fun

Now lets get some agar mixed up. Some prefer to do it in a pot with it being warmed, some do it cold. Personally I do it with warm water poured into the mix but not over a pot. Had some of my agar go jelly in the pot one time, never bothered with it since. I've got enough temperamental things in my life.

The agar recipe I used was the standard formula of 2% nutrient base 'lab standard' from Fast Freds Cookbook.

10G Malt Extract
15G Agar
1L Water

For myself, I used 300 ml of the agar for this project (the picture it shows just above 500ml with displacement of the powders the rest of the agar was used elsewhere) which makes the recipe cut in half. What you need to do is decide how many your pouring and do the math. 7ml x (how many slants) = total water needed. Adjust nutrients and agar to suit. Simple no? Here is my agar & ME on the scale.



Now stir it all together.



Time for the syringe. Stir up your agar mix to get all the powders that aren't dissolved and suck up into the syringe and start to fill your slants with maximum 7ml a piece. This is simple enough. If you happen to bugger off and come back a few minutes later, make sure to shake your syringe around to get the powders sloshed about to make an even mix for each slant.

Again, STIR BEFORE DRAWING SYRINGE

If you have CulTubes, there are two snaps. First snap is aerobic and lets air through, press harder and its air & water tight. Just leave it loose so air can get in. They'll pop off if you don't. For Vac-Tainers see attached picture. For Pyrex screw tops, just leave it slightly unscrewed to let air through. Don't worry about contams right now, just keep reading the guide, we'll get to how to avoid those.



Get a pint jar to start to lay these in. You'll fit no more then 8 using CulTubes, I think it'll be 8 with Vac-Tainers as well. Grab two lids of whatever type you decided on and lay them down then lay your pint jar down on them so you can slide the tubes in easier.



Repeat this until all tubes are filled up and all your jars are full.


Step 5: Preparing Jars for PC

Now I don't know if this is absolutely necessary but, I like to take precautions and would not want my newly poured slants to get all 'splody in my cooker while they're cooking. So I fill my jars up with some water just about to the same level as the agar in the tubes. I try not to go too high since that water may get up into the slants and throw the content off some. Your welcome to follow suit or do your own thing...don't matter to me. I'm just showing you how I get it done.





Now that all your jars are filled with a bit of water and all your tubes are capped (loosely). Now its time to put on your grain lids that you use in all your grain jars. Tyvek filters, synthetic filters, whatman custom lids, whatever...you just need some filtered G/E like you would with grain for these. This lets air pass through and around it all without needing a sterilizer to vacuum seal the whole ordeal.




Step 6: PC & Cooling

Toss them all into the PC now. I do it for 15-20 minutes at 15 PSI which is suffice for agar. I fill my PC to match the height of the water in the jars to help slow down the cooling in the jars and slants. This helps reduce or eliminate any boil over you might find. Once they are all PC'd and the PC cools enough to open it up, take your jars up to the Lab/Room with your Hood or Glove Box.

Treat these jars like you would with grain jars. No different, keep it all clean and good. Now, open your jars up and when you open up the jars the first thing you do with the slant you pull out is to either snap the lid down tight (CulTube/Vac-Tainer) or screw the lid tight (Pyrex/PP Screw top). This way you stop contams from getting in while they cool. Then lay them down so your wood stick is on the bottom of the slanted agar and let them cool in that angle till your ready to go. They will stay sealed and contam free for a long long time at room temperature. I still have slants that are in VacTainers that I didn't use and are still sitting in my drawer clean and problem free over a year later. On the note of vactainers, when your inoculating them, be sure to keep the vent hole up. Its kind of a pain, but its absolutely necessary (with either glad-wrap or parafilm, both work). But you gotta have the vent up.





And there you have it. Slants to use for your precious cultures or for precious isolates of cultures your studying and working with. You don't even need petri dishes to transfer cultures to these (although it is highly recommended). On a side note, I did have a couple of my slants have half the water/mix in them then they did when it started. It happens every time I work with these and its only 3 or 4 (even within a jar of other full well made ones). So if it happens to you, don't be bummed out. One thing for the nay-sayers out there for the method of the agar, as I finished typing up this guide I went to check my tubes, all jelled and hardened nicely as they always have without heat treatement or 'heavy mixing' like some folks say you have to do.

I hope you enjoyed this and that it will suit you well for many many years. :sun:

Edited by Mephistophelian (02/11/12 03:39 PM)

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Re: Mephisto's Master Slant Tek [Re: Mephistophelian] * 1
    #12968513 - 07/28/10 07:05 PM (13 years, 9 months ago)

Nice writeup :thumbup: Bookmarked.


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Re: Mephisto's Master Slant Tek [Re: FooMan]
    #12968779 - 07/28/10 07:59 PM (13 years, 9 months ago)

well done  :congrats: .  Your caps changed colors :wink:

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Re: Mephisto's Master Slant Tek [Re: FooMan]
    #12968846 - 07/28/10 08:11 PM (13 years, 9 months ago)

Quote:

FooMan said:
Nice writeup :thumbup: Bookmarked.



Same here. Thank you for your time and energy, you may have jsut opened my mind.

heres a quick one.. what size vacutainers? 10ml?

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Re: Mephisto's Master Slant Tek [Re: Breakfast Crew]
    #12968852 - 07/28/10 08:11 PM (13 years, 9 months ago)

Very nice tek...

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #12969114 - 07/28/10 08:59 PM (13 years, 9 months ago)

Thanks for the support guys. :grin:

I edited the Tek to answer your question Base Icks but yes, Vac-Tainers are 10ml and 16mm x 100mm where the CulTubes I used are 17mm x 100mm and PP not Pyrex (which drastically changes cost).


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #12969565 - 07/28/10 10:21 PM (13 years, 9 months ago)

Wish I had read this weeks ago when I first started playing with culture slants. Very nicely done sir, I now know what to do for next time.:thumbup:

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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #12969579 - 07/28/10 10:23 PM (13 years, 9 months ago)

Like OMG!  This is just what I was looking for.  :super:
HL


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #12970750 - 07/29/10 07:51 AM (13 years, 9 months ago)

Morning bump.  Yup.  :yesnod:
HL


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #12970809 - 07/29/10 08:26 AM (13 years, 9 months ago)

sweet tek Mephistophelian, i just started doing slants to save my cultures, been to lazy before, strange thing is some come out perfect and the odd few just dont set right, i.e like wet jelly and no good to use/

Not sure if this cause im using small ammounts i.e PDA mix for only 125 ml water usually using premade mix,

I mix up real good using off boiled water before PC'ing.

Not sure what Im doing wrong?

Is it worth giving them a little shake once out the pc to make sure its mixed properly?


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Edited by FractalXplora (07/29/10 08:26 AM)

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OfflineMephistophelian
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Re: Mephisto's Master Slant Tek [Re: FractalXplora]
    #12970866 - 07/29/10 09:03 AM (13 years, 9 months ago)

Well the one thing you want to make absolute sure of is that your calculations are right (by that I mean you have the proper nute/agar ratio to water).

I think that there is the chance that your powders weren't mixed entirely before pouring into each slant. Each time I've done these like this, I've always made sure not to let my powders get too settled before putting into each. That being said, when doing larger batches, there is always a chance that you get the odd one that exploded though I've never had one not set. Occasionally I'll get some that are cloudy and others clear.

I think next time around...stir/mix your agar slurry very well before drawing the syringe or pouring into the funnel, but personally I would think that if you do pouring there is a higher chance of getting a poor mix of powders where as with a syringe you can draw it from the bottom/center while its all still swirling and agitated. That's just IMHO.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #12970953 - 07/29/10 09:42 AM (13 years, 9 months ago)

I've had trouble with boil-over when PCing slants.  Naturally I started out loosening the lids, but I've had a whole rack of slants end up empty that way.  Tightening the lids seems to stop it, but it just seems like a dumb thing to do.  You reckon it's because they're cooling down too quickly?  I let them cool naturally in the PC..


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #12970961 - 07/29/10 09:44 AM (13 years, 9 months ago)

Nice teknology, also with pouring youre much more likely to make a mess. Do you use these for your long haul cultures?

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Re: Mephisto's Master Slant Tek [Re: badman]
    #12970981 - 07/29/10 09:53 AM (13 years, 9 months ago)

Cheers for the advice Mephisto.

I've just ordered a 100ml measuring clylinder as i been using a crappy kitchen jug. I like the idea of using the syringe to fill the slants, giving it an extra shake. Hopefully this will iron out my problems.


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Re: Mephisto's Master Slant Tek [Re: FractalXplora]
    #12971027 - 07/29/10 10:04 AM (13 years, 9 months ago)

@Andy:

The water filled inside the pint jars to provide a 'heat jacket' on the slants is what will help substantially with the cooling end of it. Just like when you heat a bottle/flask of agar, you need the water to sit 2/3 (at least) as high as your agar to reduce the cooling time as agar is extremely volatile. Even taking off the weight before your steam has finished venting (even though the PSI gauge has dropped off) can make a few explode. Using a rack (I don't own one so I'm just going out on a limb here) but I have seen them, and I don't have a clue how your supposed to have a water jacket around them to cool them down slower unless you just screw the lid on real tightly (kinda scary :shiftyeyes:).

@Badman:

I use these yes, the last handful I had used up was Vac-Tainers but I've moved to these as they are cheaper and easier to use the lids (Vac-tainers are a PITA for they're lids and very temperamental as you can see in the picture for the G/E hold). I'll take a picture of all my slants at lunch so you can see the different varieties. I plan on using Sterile water with tissue submerged with 0 GE (not to be mistaken with LC or Grain LC as those both still have nutrients in the water) as experimental for room temperature storage as well, though I would recommend against banking everything on that method since some species will die using that.

Edited by Mephistophelian (07/29/10 10:10 AM)

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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #12971274 - 07/29/10 10:57 AM (13 years, 9 months ago)

You gonna scratch a plate or submerge wedges in Eppendorf tubes? I checked up on some of my wedges in tubes and few of them have dried up but water has collected in the bottom most of them are fine. Im gonna start adding SDW to all now not only my own stock better in the long run.

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Re: Mephisto's Master Slant Tek [Re: badman]
    #12971572 - 07/29/10 12:15 PM (13 years, 9 months ago)

:yesnod:

:scaryshroom::thumbup:

:respect:


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Re: Mephisto's Master Slant Tek [Re: 13shrooms]
    #12971634 - 07/29/10 12:26 PM (13 years, 9 months ago)

Very nice Meph! Very detailed and the pics are really helpful.

Two questions- Does the shape of the wood really matter? I've been using a few long round 2mm dowels.

And- How many slants of each culture do you like to keep around in storage?

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Re: Mephisto's Master Slant Tek [Re: lazy bones]
    #12971847 - 07/29/10 01:07 PM (13 years, 9 months ago)

Great writeup.  Even took the time to include pictures.  Great work. :mushroom2: ~P~


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Re: Mephisto's Master Slant Tek [Re: lazy bones]
    #12972474 - 07/29/10 02:55 PM (13 years, 9 months ago)

Quote:

lazy bones said:
Very nice Meph! Very detailed and the pics are really helpful.

Two questions- Does the shape of the wood really matter? I've been using a few long round 2mm dowels.

And- How many slants of each culture do you like to keep around in storage?




Wood shape isn't terribly significant, as long as it doesn't severely hinder your agar slant shape and the like. (you know, like it being 1/2 wood 1/2 agar or something odd)

I actually keep 4 copies of each...I plan on making copies that 'library' and keeping it in another fridge off-site of my house/yard if anything were to happen all is not lost. I haven't gotten there yet since its such a massive under taking but...soon. Very soon.


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Re: Mephisto's Master Slant Tek [Re: FooMan]
    #12973547 - 07/29/10 06:03 PM (13 years, 9 months ago)

Quote:

FooMan said:
Nice writeup :thumbup: Bookmarked.



:rockon:


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Re: Mephisto's Master Slant Tek [Re: badman]
    #12977869 - 07/30/10 05:21 PM (13 years, 9 months ago)

Quote:

badman said:
You gonna scratch a plate or submerge wedges in Eppendorf tubes? I checked up on some of my wedges in tubes and few of them have dried up but water has collected in the bottom most of them are fine. Im gonna start adding SDW to all now not only my own stock better in the long run.




^ was a bit over my head. I think its in refference to keeping cultures in a dried stasis?

Plus this needs a :bump:

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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #12978496 - 07/30/10 07:57 PM (13 years, 9 months ago)

I'm not sure what he meant by the dried part...but you don't want to store dried mycelium. That's bad news.

What I was mentioning and he was adding to was storage of mycelium in sterile water. Its a different kind of storage system which apparently works better for some species (but will kill others so its a trade off). When doing that, you want to have it nutrient free (ie, no agar wedge, no Grain LC style mycelium etc). Just scrape the tissue off the surface of a plate and drop it into a sterile distilled water filled vial.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #12978588 - 07/30/10 08:21 PM (13 years, 9 months ago)

Ah right, thanks for the info. how do I find out what strains do well with that style?

Also I was just reading the other day that commercial strains are kept dry, and that that is the common way for commercial grow ops to maintain Iso's. Says they grind the whole fruit dried fruit and powder it. :eek:

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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #12978607 - 07/30/10 08:24 PM (13 years, 9 months ago)

I'd be quite curious to know where you found that information. Every place I have dealt with keep they're cultures in either Nitrogen Cryogenics or refrigeration. Never dried fruits. That would be the absolute worst way to keep a mother culture. You'd want that culture stored as close to its original set of genetics as possible and kept in a form that was easily accessible. A dried fruit body would be old, dry, inaccessible and...:facepalm: Just....no.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #12978692 - 07/30/10 08:48 PM (13 years, 9 months ago)

Here you go man,

Look out RR gets knee deep in there.

PS somewhere in there is a link that goes back to an even older thread.

Talk about the land before time :wink:

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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #13040330 - 08/12/10 07:14 AM (13 years, 9 months ago)

I tried RR's method of washing  chunks of WELL dried cyan giving it the 4 step sterile water bath in jars as described in his tek, I was amazed did actually get a clean culture out of about 5 chunks i stuck on agar.

Never grew em out , but I was totally amazed at the power of mycelium and the fact the that oyu could get living mycillum form a super dry old shroom.


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Re: Mephisto's Master Slant Tek [Re: FractalXplora]
    #13040405 - 08/12/10 08:06 AM (13 years, 9 months ago)

link to that tek?

:thumbup:

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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13040516 - 08/12/10 09:09 AM (13 years, 9 months ago)

Mephistophelian,


In your experience,


since you said these were no good,
http://themadseason.com/index.php?main_page=product_info&cPath=6&products_id=28


I was wondering if these would make viable tubes for culture slanting?


 





TO me they kinda look like the ones you're using


but


I'm not sure so I though I would ask.


HL


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13040521 - 08/12/10 09:11 AM (13 years, 9 months ago)

Those look like they are sealed tops possibly with a rubber/silicone top no?

Unless that cap is removable and able to allow for GE won't be any good.

Now, the madseasons Vac-Tainers do work...but they are the last option you'd go with out of all of them from my experience. And I've prepared all 100 of them and used 77 of them. Not the best option but they do work in a pinch.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13040565 - 08/12/10 09:26 AM (13 years, 9 months ago)

Quote:

Those look like they are sealed tops possibly with a rubber/silicone top no?




Sealed tops?  No.





The caps are rubber with injector port in the center.





Just wrap the where the cap meets the tube with parafilm


once prepared and inoculated?


HL


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13040579 - 08/12/10 09:30 AM (13 years, 9 months ago)

Yes, I suppose that would work. It will be cumbersome and a PITA but they will work.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13040609 - 08/12/10 09:40 AM (13 years, 9 months ago)

:strokebeard2:


Hmmm.  I ask for educational purposes.


I have never dabbled in master culture slantism.


Check out what I have going down for that one thing we've been hackling about.








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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13040618 - 08/12/10 09:43 AM (13 years, 9 months ago)

Wonderful! I actually found a clone laying around...I'm going to get it clean and see if its still ready to fly. If it is, it will cut down time considerably.

Also, word to the wise...if your going to try dabbling with a PITA like those vials, just do 1 pint jars worth of vials to test with. See if you want to put more effort in beyond that then. If not, hit me up for some of these cultubes, they're cheap.


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13040646 - 08/12/10 09:52 AM (13 years, 9 months ago)

Quote:

I have never dabbled in master culture slantism.




Me too man, these are my second attempts this time with stick for long term storage,i realized my last lot didnt have  a large enough slant / surface area,13ml in with stick seemed right. Followed Hamloafs PDA tek % 8 made just the right amount of the PDA, and slanted up via Mephistophelian tek to the letter - the grain lids / tyvek. Cheers guys have to say the shaking of and filling with the syringe worked a treat, important step no doubt to getting a nice even mix and a proper set. Done 13 all came out tops! Killer teks!

Teks in my sig now;)





:rockon:  :toast:  :headbang:


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Re: Mephisto's Master Slant Tek [Re: FractalXplora]
    #13040676 - 08/12/10 10:02 AM (13 years, 9 months ago)

Beautiful! Great that it works so well for you! Really like seeing results and helping people out with what seems like such a huge problem and turning into something manageable. :awesome:


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13040712 - 08/12/10 10:13 AM (13 years, 9 months ago)

I felt i needed to learn to make em, been givin some great strains this summer from kind folk on here and just aint got the time so i need long term storage to do them justice. Many thanks guys.:thumbup:


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Re: Mephisto's Master Slant Tek [Re: FractalXplora]
    #13041050 - 08/12/10 11:42 AM (13 years, 9 months ago)

Nice write up. Thank you :smile:


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Re: Mephisto's Master Slant Tek [Re: Jitsu]
    #13050309 - 08/14/10 11:53 AM (13 years, 9 months ago)

I don't even have a 15psi cooker yet (only a 10) but read this out of curiosity and I have a question. Your tubes look like they go up to 10ml. In step 2 it says to add a stick and then water to 5-7ml but then in step 4 it says to add 7ml of the agar solution.

When does the 5-7ml of water get removed? Why do you use it? What happens to the sticks from step 2? It looks like there is only one stick in each tube.

Thank you for posting this. I hadn't read about how to do anything with agar before but I think I'll try this when get a better PC.

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Re: Mephisto's Master Slant Tek [Re: sve]
    #13051140 - 08/14/10 03:17 PM (13 years, 9 months ago)

The reason I did the water thing and laying it down was so the person who was working with (who would probably have a different type of slant then I used in the example) could see if 7ml was enough to make the right amount of surface area when it was on a slant. That and you need to get your 'slanted surface' setup in your clean area before you get the agar cooked. You don't want to try to guess while the agar is hot and ready to cool. Once you've figured out that 7ml was suffice, you dump the water out but you keep the sticks. You only need 1 stick per tube.

10PSI pressure cooker will still work for this slant method. You simply add an extra 10-15 minutes to your sterilization cycle to compensate.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13051183 - 08/14/10 03:32 PM (13 years, 9 months ago)

Thanks, that makes a lot more sense now! Are those tubes the kind that the chemistry department at my college might have?

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Re: Mephisto's Master Slant Tek [Re: sve]
    #13051209 - 08/14/10 03:41 PM (13 years, 9 months ago)

Perhaps. If they do any culture work of any kind, there is a really good chance they'll have these. If not these then they would have the expensive Pyrex with screw top types.

But I have empty and pre-poured ones available in case anyone can't get a decent/cheap source of slants.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13051219 - 08/14/10 03:44 PM (13 years, 9 months ago)

I have no idea what they do there, to be honest. Would a biology department be more likely to use them?

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Re: Mephisto's Master Slant Tek [Re: sve]
    #13051242 - 08/14/10 03:52 PM (13 years, 9 months ago)

Yes, if anything you'd talk to Biology about getting some autoclavable tubes. At the minimum PP (Polypropylene) and the expensive option Pyrex.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13051348 - 08/14/10 04:18 PM (13 years, 9 months ago)

First off, nice write up.  This is something I've recently become interested in.

One n00by question:  What is the purpose of having a wood stick in the slant?


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Re: Mephisto's Master Slant Tek [Re: BrainFarmer]
    #13051382 - 08/14/10 04:28 PM (13 years, 9 months ago)

Thanks!

The stick is a hardwood nutrient/matter for the mycelium to get a really solid hold onto instead of just pure nutrient agar mix. From what I understand, this ensures a longer life and better rejuvenation for the mycelium down the road.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13051450 - 08/14/10 04:45 PM (13 years, 9 months ago)

Let's talk about master culture slants, shall we?  The wood stick is for a for an extra food source yes.  The myce could not live for 2 years, in the fridge, off of just agar alone.  The low temperatures of the  fridge, slows down the metabolic rate of the myce.  Couple this with the extra food source (wood stick) and myce will store for a couple years in the fridge.  Master culture slants, pure genius.
LM


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13106222 - 08/26/10 04:42 PM (13 years, 8 months ago)

Nice Mephisto!
tiny tip! throw a quarter of a sterile tooth pick in a LC or onto an isolated sector on a plate. give it some time to grow over then pick it out with sterile tweezers, throw it into the master slant to get it going.

Beats the hell out of trying to cut a wedge out...trying to geT IT OFF THE BLADE!!! did I touch the lip? was that sterile? I think I stuck my finger in the end of the slant....DROPPED IT!!!! FUUUUUUUUUUUUUU!!!!!! WHERE IS THE LID!!!

works the other way too...put the sterile tooth pick in the master slant then move it to agar then grow it out.


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13106621 - 08/26/10 06:42 PM (13 years, 8 months ago)

I just did my first plate transfers from slants today. I noticed how hard it is to get wedges off of the scalpel. What a ball ache.

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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #13106643 - 08/26/10 06:49 PM (13 years, 8 months ago)

Quote:

Base Icks said:
I just did my first plate transfers from slants today. I noticed how hard it is to get wedges off of the scalpel. What a ball ache.



I just use the tip of a syringe as a tiny scalpel, works great and you can spear it when it's free.
and there is always one nearby, they are all sterile...can't miss. works great for clones too.


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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #13106710 - 08/26/10 07:07 PM (13 years, 8 months ago)

Quote:

Base Icks said:
I just did my first plate transfers from slants today. I noticed how hard it is to get wedges off of the scalpel. What a ball ache.




They're sticky buggers.  If it won't come off easily in a face-up manner, I've found it doesn't seem to hurt to put them face-down -- i.e. turn it upside-down and stick the mycelium side to the receiving dish and slide the scalpel away.  The fungus, she don't seem to mind at all.


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Re: Mephisto's Master Slant Tek [Re: andymc]
    #13108647 - 08/27/10 05:26 AM (13 years, 8 months ago)

Sorry for the of topicness.

I've also noticed that its a bitch to get my scalpel red hot. Seams like the very tip gets hot but thats it. I'm using a regular lighter on a proper scalpel.

The lighter brings me to another thing. All the soot on the blade from flaming came off in my slants when I was getting wedges out of them. Is that a problem?

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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #13108659 - 08/27/10 05:33 AM (13 years, 8 months ago)

The lighter is causing the soot problem.  Hardware and camping stores sell aerosol aerosol-like cans with propane/butane, and reusable nozzles that you screw on top.  They can get a syringe glowing in seconds, and they don't leave much soot
Honestly, I don't flame my scalpel any more (bad andymc! :punish2:).  I disinfect it with isopropyl before each dish.  I never get any obvious contams from taking that little shortcut (i.e. no contams starting around the wedge).


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Edited by andymc (08/27/10 05:35 AM)

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Re: Mephisto's Master Slant Tek [Re: andymc]
    #13108762 - 08/27/10 06:23 AM (13 years, 8 months ago)

You guys need to be getting propane torches. Work amazing! Nothing gets that blade hotter and faster then a propane torch.

And they're cheap :yesnod:


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13108836 - 08/27/10 07:04 AM (13 years, 8 months ago)

Quote:

Mephistophelian said:
You guys need to be getting propane torches. Work amazing! Nothing gets that blade hotter and faster then a propane torch.

And they're cheap :yesnod:



:hammertime:


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13108851 - 08/27/10 07:11 AM (13 years, 8 months ago)

I'm going to get a torch lighter. Acceptable?

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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #13108887 - 08/27/10 07:22 AM (13 years, 8 months ago)

Quote:

Base Icks said:
I'm going to get a torch lighter. Acceptable?




How about this?


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Re: Mephisto's Master Slant Tek [Re: andymc]
    #13108905 - 08/27/10 07:28 AM (13 years, 8 months ago)

Quote:

andymc said:
Quote:

Base Icks said:
I'm going to get a torch lighter. Acceptable?




How about this?




Both acceptable, though in terms of longevity, Andy's is a better choice.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13108912 - 08/27/10 07:32 AM (13 years, 8 months ago)

I get the gas canisters for mine at the local DIY store.  A big tall can (about double the size shown) has lasted me 6 months so far..


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Re: Mephisto's Master Slant Tek [Re: andymc]
    #13108916 - 08/27/10 07:36 AM (13 years, 8 months ago)

Ah, you already have one. I understand now :yesnod: And yes, they last a really long time. Lasts me about the same amount of time. Safer then a little glass lamp too. Worst case you have a pointed flame laying somewhere, not a glass jar full of liquid alcohol waiting to make an instant moltov cocktail.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13110287 - 08/27/10 02:25 PM (13 years, 8 months ago)

This is the torch I bout from home depot and the sell small bottles of fluid for it.  Works wonders..

http://www.google.com/products/catalog?hl=en&q=bernzomatic&cid=895655214206422099&ei=KR94TN7DHIO62QTJtbz8Dw&sa=title&ved=0CB8Q8wIwAjgA#p

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13110290 - 08/27/10 02:26 PM (13 years, 8 months ago)

It has a nice little button that once lit will lock it in the on position so you don't have to hold it down so you can lock it on then leave it sit there while you work and not worry about starting it back up.

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13110764 - 08/27/10 04:12 PM (13 years, 8 months ago)

I actually have one of those that andy posted. Its got a really tall can probably like what andy has. Even got an electronic start.

I dont know why I hadnt thought about that. I guess it just struck me as overkill.  :minigun: :zombie2:  :machinegun:

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Re: Mephisto's Master Slant Tek [Re: Base Icks]
    #13110887 - 08/27/10 04:40 PM (13 years, 8 months ago)

I just happen to find what I have.

Thanks Step Dad.

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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13110926 - 08/27/10 04:53 PM (13 years, 8 months ago)

Quote:

Mephistophelian said:
You guys need to be getting propane torches. Work amazing! Nothing gets that blade hotter and faster then a propane torch.

And they're cheap :yesnod:



LOL I taught you that one I think!


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13110942 - 08/27/10 04:59 PM (13 years, 8 months ago)

Nope, I did.
LM


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111040 - 08/27/10 05:39 PM (13 years, 8 months ago)

You buncha whack jobs :lol:

I grabbed a torch because my alcohol burner cracked in 6 places and turned into a "fires of Kuwait" while I was working.

P.S. I'm the true inventor of Tin Foil :awesome:


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111076 - 08/27/10 05:52 PM (13 years, 8 months ago)

King Size!


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13111096 - 08/27/10 06:01 PM (13 years, 8 months ago)

You ain't got to lie to kick it. 

You grabbed that torch because I suggested it to you that time you came crying to me.
Talking about how your alcohol lamp broke because you were weren't using it right.
You needed a replacement but didn't want to buy another alchol lamp.

You were becoming wise in the ways of Mycology by this point.
You began to see the design flaws of an alcohol lamp.
As your mentor, that's when I realized you were ready to be introduced to the torch as a replacement to the alcohol lamp.
Which at the time, you couldn't even operate correctly.
When I came through (with what seemed to your young apprentice mind at the time) the clarity and suggested you buy a propane, butane, or mapp gas torch.

All of which I, in fact, invented.
LM


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111123 - 08/27/10 06:08 PM (13 years, 8 months ago)

Says the guy who joined far later then I did no less. :lol:


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13111149 - 08/27/10 06:18 PM (13 years, 8 months ago)

He went back in time, taught your younger self in September 2008 to use the torch then came to 2009 just before Christmas and joined the Shroomery.

You don't remember because it was a time paradox.

I rest my case.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13111154 - 08/27/10 06:19 PM (13 years, 8 months ago)

Oh yeah.

Because when I joined the shroomery dictates when I began mycology.
Glad we cleared the air on that one.

Said the guy who just scored 2 AA X25 120 volt Pressure Sterilizers on ebay for under 400 dollars as well as invented the torch.
LM


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111165 - 08/27/10 06:23 PM (13 years, 8 months ago)

Time pardox huh !! :lmafo:

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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111166 - 08/27/10 06:24 PM (13 years, 8 months ago)

Quote:

LiquidMyce said:
Oh yeah.

Because when I joined the shroomery dictates when I began mycology.
Glad we cleared the air on that one.

Said the guy who just scored 2 AA X25 120 volt Pressure Sterilizers on ebay for under 400 dollars as well as invented the torch.
LM




How do you get 120 volts out of 2 AA batteries?

And why would you buy 25 of them?


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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13111170 - 08/27/10 06:26 PM (13 years, 8 months ago)

I WILL HAVE THIS THREAD AT 1000 VIEWS BY THE END OF THE NIGHT!
All hail ME!
LM


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13111178 - 08/27/10 06:29 PM (13 years, 8 months ago)

Quote:

Peterthinks said:
He went back in time, taught your younger self in September 2008 to use the torch then came to 2009 just before Christmas and joined the Shroomery.

You don't remember because it was a time paradox.

I rest my case.



Watch it Peterthinks, are you sure about that?

I am 47 years old.
In human years, that translates to 1002 years old.
LM


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111185 - 08/27/10 06:31 PM (13 years, 8 months ago)

Your an old fucker !!

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13111191 - 08/27/10 06:34 PM (13 years, 8 months ago)

:oldman:


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111197 - 08/27/10 06:36 PM (13 years, 8 months ago)

LoL question for ya shouldn't u be collecting social security and not growing mushrooms.  :laugh2:

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13111255 - 08/27/10 06:54 PM (13 years, 8 months ago)

Quote:

Damion5050 said:
LoL question for ya shouldn't u be collecting social security and not growing mushrooms.  :laugh2:



Actually, there can be no gauge on my age because I am immortal.
We do not require this "Social Security" you speak of, as well.

Also,

Furthermore, I do not "grow shrooms."
"Growing shrooms" is for mere mortal human beings.
I encourage mushrooms to grow in their natural state of mushroomness, Mush Doctor.
LM


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111264 - 08/27/10 06:57 PM (13 years, 8 months ago)

LoL whatever..  Who are you Craig Christ.  Jesus's long lost brother ??

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13111286 - 08/27/10 07:08 PM (13 years, 8 months ago)

Quote:

Damion5050 said:
Your an old fucker !!




Hey,  47 is a very cool age, I'll have you know!

;0)

JD


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Re: Mephisto's Master Slant Tek [Re: Javadog]
    #13111294 - 08/27/10 07:11 PM (13 years, 8 months ago)

Well other than it being a prime number I can't think of anything really cool about it.


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13111342 - 08/27/10 07:24 PM (13 years, 8 months ago)

:dumblol:

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13111380 - 08/27/10 07:33 PM (13 years, 8 months ago)

:crazybeard:


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13111389 - 08/27/10 07:35 PM (13 years, 8 months ago)

Quote:

Peterthinks said:
Well other than it being a prime number I can't think of anything really cool about it.




Ah, but it is a safe prime!


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13111424 - 08/27/10 07:41 PM (13 years, 8 months ago)

Quote:

Damion5050 said:
:dumblol:




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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111572 - 08/27/10 08:22 PM (13 years, 8 months ago)

And a part of the Fibonacci sequence but even that doesn't make it cool.


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13111608 - 08/27/10 08:33 PM (13 years, 8 months ago)

Quote:

Peterthinks said:
Quote:

LiquidMyce said:
Oh yeah.

Because when I joined the shroomery dictates when I began mycology.
Glad we cleared the air on that one.

Said the guy who just scored 2 AA X25 120 volt Pressure Sterilizers on ebay for under 400 dollars as well as invented the torch.
LM




How do you get 120 volts out of 2 AA batteries?

And why would you buy 25 of them?



Don't be so sure of yourself there, big guy.



Just one of these things can PC 2 Presto 23's all day quietly and with little energy used!
LM

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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13111636 - 08/27/10 08:40 PM (13 years, 8 months ago)

LOL

Well I may have to upgrade!

Holly crap I need a smoke....I'm gonna go make some agar, Later. I need to keep busy.


Trying to quit.


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13111652 - 08/27/10 08:44 PM (13 years, 8 months ago)

LM what model is that ??  And how much does it hold ??  I been deciding if I want to buy a 25 50 or 75x

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Re: Mephisto's Master Slant Tek [Re: Damion5050]
    #13112087 - 08/27/10 10:38 PM (13 years, 8 months ago)

Quote:

Damion5050 said:
LM what model is that ??  And how much does it hold ??  I been deciding if I want to buy a 25 50 or 75x




I think that they are the All American 925 Steroclave.
(25 quarts)

I got one off Bay for about $200.  Great deal.

I could be bigger, as you love a couple inches of depth
for the coil and stand, but it works very well.
(easy to control temp...no hissing at all)

JD


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Re: Mephisto's Master Slant Tek [Re: Javadog]
    #13112106 - 08/27/10 10:42 PM (13 years, 8 months ago)

Quote:

Javadog said:
Quote:

Damion5050 said:
LM what model is that ??  And how much does it hold ??  I been deciding if I want to buy a 25 50 or 75x





I could be bigger, as you love a couple inches of depth



JD





ANYONE???.....


Insert joke here.


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13112119 - 08/27/10 10:44 PM (13 years, 8 months ago)

MUST.....NOT.....GET......BANNED!


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13112122 - 08/27/10 10:44 PM (13 years, 8 months ago)

LOSE!

I blame weed.

;0)


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13112126 - 08/27/10 10:45 PM (13 years, 8 months ago)

GARRRUGHHHHH!!!!!


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Re: Mephisto's Master Slant Tek [Re: Javadog]
    #13112131 - 08/27/10 10:46 PM (13 years, 8 months ago)

Quote:

Javadog said:
LOSE!

I blame weed.

;0)



Beer actually, but thanks for asking.


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13112218 - 08/27/10 11:08 PM (13 years, 8 months ago)

Quote:

LiquidMyce said:
I WILL HAVE THIS THREAD AT 1000 VIEWS BY THE END OF THE NIGHT!
All hail ME!
LM



Yup, I think my work here is done.

Quote:

Damion5050 said:
LM what model is that ??  And how much does it hold ??  I been deciding if I want to buy a 25 50 or 75x




Damion, we will disscuss this and more in the morning when I start my own thread.  :super:
LM


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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13112882 - 08/28/10 04:06 AM (13 years, 8 months ago)

I wake up to 26 new post in this thread, and thats all I get?

I very disappointed in at least a few of you. :header:

PS

And now you broke the sever and I cant post this.

Probably for the better, I have a productive day ahead of me.

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Re: Mephisto's Master Slant Tek [Re: hamloaf]
    #13113160 - 08/28/10 07:36 AM (13 years, 8 months ago)

Lets get this thread back on topic shalt we? :waits:


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13116837 - 08/28/10 10:24 PM (13 years, 8 months ago)

Mephisto, do you add anything to your agar to make a master slant?
What is your agar recipe?

Back on track!!!


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Re: Mephisto's Master Slant Tek [Re: Peterthinks]
    #13117057 - 08/28/10 11:32 PM (13 years, 8 months ago)

My agar recipe is just the same as my dishes.

10g Agar
15g Malt Extract
500ml water


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13120643 - 08/29/10 07:19 PM (13 years, 8 months ago)

Quote:

Mephistophelian said:
My agar recipe is just the same as my dishes.

10g Agar
15g Malt Extract
500ml water




Hey,

I checked a recipe that I had written in my grow log, and
saw 10g LDM + 9g agar + 500 ml H2O.

Hmmmmm....I wonder...

JD

P.S. The formula that I found has worked fine for me.
(less nutritious it would seem though)


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Re: Mephisto's Master Slant Tek [Re: Javadog]
    #13120911 - 08/29/10 08:14 PM (13 years, 8 months ago)

Your recipe would work just as well.

I've always used this recipe from FF's CookBook

Quote:

Malt Extract Agar (3%)(MEA)(General Microbiology)(FDA M93)
-----------------------------------------------------------------
30 g Malt extract
20 g Agar
1 L Distilled water




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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13120973 - 08/29/10 08:26 PM (13 years, 8 months ago)

Well, Fastfred is certainly reputable.  ;0)

Carry on!

JD


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Re: Mephisto's Master Slant Tek [Re: Javadog]
    #13122903 - 08/30/10 09:07 AM (13 years, 8 months ago)

I believe 2-4% is an acceptable sugar level. :super:

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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13597383 - 12/07/10 06:05 PM (13 years, 5 months ago)



Good stuff. :thumbup:

Edit- Can someone please link me to a site that sells those pyrex screw caps in the pics above? I can't remember where I got those....I want to make MOAR!!!1 :biggrin:


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Re: Mephisto's Master Slant Tek [Re: fungus_tao]
    #13625985 - 12/13/10 11:25 AM (13 years, 5 months ago)

Thank you for the write up Mephisto, I have the PC fired up and sterilizing away..You tec was a breeze to follow..I learned a lot...peace n shrooms be upon you....:mushroom2::mushroom2::mushroom2::mushroom2::mushroom2:

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Re: Mephisto's Master Slant Tek [Re: Ozzy]
    #13640972 - 12/16/10 08:39 AM (13 years, 5 months ago)

Your very welcome Ozz...this was one tek that was never shown in extent (or at all) from my knowledge beyond what RR did in his video's which didn't cover how to do it without a rack nor any other types of slants so it needed to be done. Can't have everyone losing they're cultures due to being scared of trying a simple thing :wink:


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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #13642454 - 12/16/10 02:39 PM (13 years, 5 months ago)

I love this site and all the GREAT people that write up tek like this one. Thank Mephisto for taking the time to make this site the best one on the net.:murray:


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Re: Mephisto's Master Slant Tek [Re: pepper]
    #13725080 - 01/03/11 02:19 PM (13 years, 4 months ago)

Posting to follow this outstanding writeup  :nothingtoadd:


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Re: Mephisto's Master Slant Tek [Re: Fungicide]
    #13725363 - 01/03/11 03:22 PM (13 years, 4 months ago)

Yup, this is one of the good ones.

...and as I am about done capturing strains as prints, I will be
moving to isolation and then slants will be important.

Take care,

JD


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InvisibleHorizonSpawn
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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #15285248 - 10/27/11 09:48 AM (12 years, 6 months ago)

Necro-threading this one for good reason: WOW!  Most awesome write-up and TEK!

Thank you, thank you, thank you! :thumbup:


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Please assume any and all prints exchanged are "WILD" in nature; and thus, should NOT be considered ASEPTIC...

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OfflineMephistophelian
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Re: Mephisto's Master Slant Tek [Re: HorizonSpawn]
    #15315538 - 11/02/11 10:07 PM (12 years, 6 months ago)

Your very welcome =) I'm glad I could help you out.


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Master Slants Made Simple

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InvisibleFungus_monk
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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #15315595 - 11/02/11 10:19 PM (12 years, 6 months ago)

Hey OP,
In the bottom of your original post you say that you have pre-poured tubes ready. I realize this is over a year old but if you still have some please :pm: me cause im definitly interested in a couple.
Awesome write-up btw!


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Offlinemikesethnobotany
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Re: Mephisto's Master Slant Tek [Re: Fungus_monk]
    #15315663 - 11/02/11 10:32 PM (12 years, 6 months ago)

OP

I think that it may be nice to add in that you need to parafilm the tubes after inoculation, especially for the vacutainers. I made these and my cultures didn't grow hardly at all for a few weeks until I realized that having the red SHIP all the way down was depriving the slant of gas exchange. So I just propped it up to its gas exchange hole and wrapped that small portion with parafilm likeso:


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OfflineMephistophelian
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Re: Mephisto's Master Slant Tek [Re: Fungus_monk]
    #15315674 - 11/02/11 10:35 PM (12 years, 6 months ago)

Whoo...actually my flowhood is packed away right now, so there's no way I could do any pre-poured ones at the moment, even if I wanted to. I have tubes and supplies, just no means to cool them cleanly :/

@Mike, I did take a picture of the vac-tainer with the lifted bit. Maybe I could be more clear about the inoculation part...

Edit: Added note to main post.


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Master Slants Made Simple

Edited by Mephistophelian (11/02/11 10:38 PM)

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Offlinemikesethnobotany
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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #15315696 - 11/02/11 10:41 PM (12 years, 6 months ago)

Thanks mephisto! Great writeup, +5

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Offlinegratefulskinny
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Re: Mephisto's Master Slant Tek [Re: mikesethnobotany]
    #15316293 - 11/03/11 02:18 AM (12 years, 6 months ago)

super informative +1!!!

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OfflineMephistophelian
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Re: Mephisto's Master Slant Tek [Re: gratefulskinny]
    #15792466 - 02/11/12 12:28 PM (12 years, 3 months ago)

Just a bump for any new members that might have missed this :awesome:


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Offlinetotal
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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #15792553 - 02/11/12 12:46 PM (12 years, 3 months ago)

:fuckinawesome:

And also Mephistophelian, i would suggest a much lower Malt content...
Specially for slants...:shrug:

I personally recommend 1g ELME per 100ml dh2o

Edited by total (02/11/12 12:51 PM)

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Invisiblearis
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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #15792592 - 02/11/12 12:55 PM (12 years, 3 months ago)

What are the advantages of doing this with agar instead of distilled water?

http://www.shroomery.org/forums/showflat.php/Number/14632590#14632590

Quote:

lipa said:
Just some extra info. I always make backups with culture tubes and distilled water. It is important that the wedges be very small 3-4mm diameter and put 3-4 of them in each tube. You don't have to use cryo-vials either. I use transport tubes that are 5ml.

There is an advantage to making distilled water vials too.  If you make a few of them for each strain it is very easy to make grain spawn masters without having to get the mycelium running on agar first. Just make up some grain and drop the wedges strait into the grain and done.

I have been thinking about growing mycelium over small pieces of wood and dropping them in to distilled water as well. You could use cardboard most likely. I have also used those little kitty litter cardboard balls they sell at the pet store moistened and loaded in small tubes with mycelium. I recently pulled a three year old shiitake culture out of the fridge and placed a small chunk on agar. Took of like a rocket and looks really healthy. Agrocybe agertita stores really well this way too.

Don't expect distilled water storage to work every time. It doesn't with every species. That's why I do both so I will know which ones will work like that in the future (I also like the ability to just make up grain spawn masters without having to have an extra plate out).  Storage vials take up very little room. That's nice when you are limited on space
.
Lipa



Edited by aris (02/11/12 01:05 PM)

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Offlinetotal
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Re: Mephisto's Master Slant Tek [Re: aris]
    #15792656 - 02/11/12 01:13 PM (12 years, 3 months ago)

Distilled water storage is a 3d storage...
I prefer growth on agar b/c the culture grows on a 2d plane, it makes seeing contaminates easier...
And when your working with master/backup cultures, hidden contamination is the last thing you want...

:2cents:

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Offlinewinemaker
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Re: Mephisto's Master Slant Tek [Re: Mephistophelian]
    #26037311 - 06/06/19 09:07 PM (4 years, 11 months ago)

And today I learn what slants are. Thank you for the write-up!


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OfflineMephistophelian
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Re: Mephisto's Master Slant Tek [Re: winemaker]
    #26044779 - 06/10/19 08:54 PM (4 years, 11 months ago)

You are very welcome! Glad it could be of use even after all these years :laugh:


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