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Offlinekadmsu
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Registered: 09/30/11
Posts: 23
Last seen: 10 days, 19 hours
Two Syringes: Here's my plan w/ questions
    #23936588 - 12/17/16 09:24 PM (7 years, 2 months ago)

Ello!

Been off of my mycology game for quite some time, and was never very experienced, just read a lot. Decided it's about time to get a little more serious.

So I just got two 10 cc MS syringes and wanted to run my plan by some more eyes and ask a few questions. Here it is:

I'm going to inoculate WBS jars, 7 from 1 syringe and 8 from the other for a total of 15 Quart jars.  Not sure if that is too many jars for one syringe, any input here would be welcome.

To inoculate I am going to quickly remove the metal ring and filter disc, and inject ~ 1-1.5 cc on the side of the container, and recap as quickly as possible, as I have no injection port.

Once they colonize I will spawn two monto tubs, case when colonized, and fruit.

From then I will take a few clones of the best looking fruits and plate them on agar.

To complete the cycle I will inoculate more WBS spawn and repeat sans the cloning again.

If I clone from a fruit and use that mycelium to spawn more tubs, will that suffice for ensuring solid flushes for future monotubs? 

At some point I would love to do some strain isolation. In that case is it better to start from a clone or from spores?  I read that even if you clone from fruits you still get multiple sectors of mycelial growth.  I would worry about starting from spores and not isolating a fruiting strain.  I don't have endless supplies to select multiple strains, isolate them, and fruit them, so I am thinking isolating from a fruit would be better considering my situation?

Also, how beneficial is it to isolate a strain?  What are the benefits working with an isolated strain?  Faster colonization? More prolific and even fruiting?

Thanks in advance everyone!  I tried to do my research but I'm sure I'm asking questions that have already been asked and answered, so I apologize for that.

Cheers!

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Invisibler.lutece
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Re: Two Syringes: Here's my plan w/ questions [Re: kadmsu]
    #23936605 - 12/17/16 09:32 PM (7 years, 2 months ago)

First off: Be prepared for a lot of people to tell you that you shouldn't go spore syringe to grains; it has a higher failure rate than other methods, so a lot of people knock it in favor of doing spores to agar from the get-go.

  • That's a pretty good number of jars for each syringe for grains.

  • Are you sterilizing jars? They'll need to be pressure cooked at 15psi for 90-120 minutes. (Steam sterilization rarely cuts it.)

  • Are you inoculating inside of a still air box? If you do this open air, your chances of contamination go way up once you open your sterilized jars.

  • If you clone to agar, I wouldn't bother noc'ing with syringes the second time around; that's where your agar should come in. (Not sure from the wording if that's what was planned.)

  • Why not clone on the second grow?

  • If you clone a fruit, it will replicate that fruit's tendencies. (For example, if you clone a fruit from a cluster, you'll get more clusters. That's a good way to get a bunch of fruits in a harvest.)

  • Isolating from a clone is faster; there are fewer strains to isolate away from when starting from a cloned fruit versus starting with spores. You'll likely have to do fewer isolations because of this. That being said, not all strains inside of a fruit will be fruiting strains, so remember to test each isolation by growing it out small-scale before taking it to bulk.


I am also interested in knowing the specific advantages of working with an isolation.


--------------------
One goes into an experiment knowing one might fail.
But one does not undertake an experiment knowing one HAS failed.

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OfflineTh3Issu3
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Re: Two Syringes: Here's my plan w/ questions [Re: kadmsu]
    #23936619 - 12/17/16 09:37 PM (7 years, 2 months ago)

First, do you have a SAB and pressure cooker...

The most important thing in this whole process is that everything is  steril. That's the most important thing. The tubs will come later in this whole ordeal.

You can go ms syringe to grain, but if you have agar go to agar first. And only use a few drops on a plate. That syringe can last you 300 mono tubs of you do it right.


Quote:

Th3Issu3 said:
I love seeing this everyday.


To be honest, I feel like it all boils down to how far you want to take this hobby.


I went years going ms syringe>grain. I also went years without a flow hood. In that time I had some awesome grows, full canopy's and tub after tub. I did tons of jars and bags and worked my ass off in my SAB. I had contams, everyone does. It was like shooting dice. I just bought more grain and more syringes and trucked forward.

Over last summer I took a break, I suggest everyone who's ever been a slave to there PC to take one a year. This year when I came back at it, I told myself I was going to take it to the next level.

I bought a flowhood. Told myself I was going to get into this agar hype all the older guys and Tc's always swore by. So that's what I did. Got some already mixed potato dextrose agar. The simplest kind I could find. Just add water.

..Now at this time I was so used to blowing through 5-10 syringes a month and having so many contam issues I went out and bought 10 gt syringes and 10 Bplus.


It's been three months after I started working with agar and with my flowhood. I have 9 of each of the needles left. Have had basically no contams and im rocking and rolling at a 100 % contam free rate on all my jars and tubs. I'm saving money on syringes b/c its only taken one drop to make a plate which i clean up then make 4 more plates, then tiger drop into masters then grain to grain. Saving tons of money on grain, and there is less clean up.

Basically for me, It's now a no brainer. Can you go from ms syringe to grain.... Sure, why the fuck not. Will you save time, money, and heartace going to agar first? fuck yeah dude!

It's all boils down to how far you want to take this hobby my friend.

Good luck





Also read this.....


Quote:

Th3Issu3 said:
When working with agar, my goal is clean healthy growth. I'm not trying to isolate anything. So the amount of transfers I do solely depends on that. Each time I transfer from my M.S. plate I also label it "T" then the number, for example; T1,T2,T3,ect... Usually 'T-2' Is where I find myself before cutting the plate up and using it for inoculation.

My process in this, is in steps like this......

MS to petri, labeled 'ms p'
'ms p' to 3 new plates labeled 'T 1'
3, 'T 1' plates to 3, 'T 2 ' plates

*all plates are saved for a week or so in case i need to use them again. But after clean growth on the donor plat is achieved I toss them.

Now depending how the plates look 't-2' is where I start using wedges. Not always... But like i said, depending if they are clean on not. 

So I now have 3, 't-2' petri's with clean growth. 

I make 16; 1 pint master.

I now take plate 1 and 2 labeled "T-2" and cut it up and use them to innoc the 16 1 pint grain masters. I do this b/c I take the 1 pint grain master after 100 % colonization to inoculate 2, 2 quart grain bags. In which I just use half of the jar in each bag. That's what I use for a tub. About 4 quarts of spawn...

OK now you have one plate left...

First thing i do is make 3 slants. I do this so i dont have to start from scratch...

So I take a small pieces from my t2 agar plate and drop them in the slant and label them T-3' then the strain and date. Those get set aside to fully colonize. Once they are, they get put in a lunch box in the fridge. ( I will will use this in the up coming months)

You still have the 't-2' pate just a few chunks missing...So what do we do...More transfers... Now following what I do here you can keep going down the line as far as you want.

I take the "T-2" petri and make 3 plates. Labeled "T-3"...Then I just do the same thing I did previously. Take two plates for masters then use one to go further down the line.

I try not to go farther than 't-6'. I'm not trying to isolate genetics. Just want clean growth.

When you transfer out of t-6 bring your slant out and do the same thing, work up to t-6.

IDK how others do it, but this has worked well for me. Wish you the best!!





Also people isolate to find genitics that benefit them..

Fast growth, potency, full canopys.....things of that nature.

Edited by Th3Issu3 (12/17/16 10:02 PM)

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Offlinekadmsu
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Registered: 09/30/11
Posts: 23
Last seen: 10 days, 19 hours
Re: Two Syringes: Here's my plan w/ questions [Re: Th3Issu3]
    #23936709 - 12/17/16 10:11 PM (7 years, 2 months ago)

Thanks for the quick and informative responses r.lutece and Th3Issu3!

I have both a steroclave and a flow hood so hopefully that will reduce my contamination events.  Even with these is grain still difficult to successfully inoculate with spores?  From Th3Issu3 post it seems like going agar first is the way to go.


r.lutece for the 2nd go around I was going to use the agar wedges from my cloning to inoculate; wasn't too clear about that.  As for cloning on the 2nd round, I figured I would just keep doing g2g for a while.  I know at some point the mycelium starts to senesce, but I figured I could get a few tubs before cloning again.


Th3Issu3 that is some good info.  Sounds like a good system for tracking mycelial expansion.  I definitley want to do some slants to so I can preserve good genetics.

Ive got a decent understanding of all these techniques, but have never actually done them so I am a bit nervous.

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OfflineTh3Issu3
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Registered: 12/30/14
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Re: Two Syringes: Here's my plan w/ questions [Re: kadmsu]
    #23936727 - 12/17/16 10:14 PM (7 years, 2 months ago)

Just do it, who cares if you fuck up. You will more than likely fuck up anyways.  I fuck everything up. That's how you learn

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InvisibleComebackKid
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Re: Two Syringes: Here's my plan w/ questions [Re: Th3Issu3]
    #23936919 - 12/17/16 11:22 PM (7 years, 2 months ago)

Dude 1cc of solution is way too much for each jar. Try 1 or 2 drops in the center of the jar. Don't shake em.
Less solution you use the better chances you have of clean spawn in the end. Considering you won't be doing agar first you want to try and make it as clean as possible still


--------------------
:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself

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Invisibler.lutece
gave Columbia her wings.
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Re: Two Syringes: Here's my plan w/ questions [Re: Th3Issu3]
    #23936924 - 12/17/16 11:25 PM (7 years, 2 months ago)

Quote:

Th3Issu3 said:
Just do it, who cares if you fuck up. You will more than likely fuck up anyways.  I fuck everything up. That's how you learn




:cantarguewiththat:

It's not so much that spores to grain will have difficulty germinating (though it can take an extra week or so in some cases); rather, because spore syringes frequently carry at least a small number of contaminants, those contams also have a very good chance of germinating, and they will frequently outrun the mycelium when it comes to colonizing the jar. That said, some syringes are quite clean, and you'll have at some success going straight to grains. The only reason folks advise against it is because it means disregarding a very common source of contamination that can be eliminated by going to agar first. But, it can definitely be done. Many of us used spores to grain in our first grows and came out the other end with shrooms to show for it, so if you want to try it out to get yourself going quickly, you'll likely still see results. You just might not get the harvest that you may have gotten with clean spawn (i.e., spawn inoculated and colonized in sterile conditions from a clean source).

As for cloning, that's just a preference thing. I believe the amount of colonization required for successful G2G results in senescence faster than agar transfers, but cloning the same line of mycelium will have more or less the same effect. I was just curious. :smile:

At the end of the day, listen to Th3Issu3: just do it. You learn a lot more from failure than you do from success, and if you succeed, you have fruits to show for your labor.


--------------------
One goes into an experiment knowing one might fail.
But one does not undertake an experiment knowing one HAS failed.

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InvisibleComebackKid
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Re: Two Syringes: Here's my plan w/ questions [Re: r.lutece]
    #23936935 - 12/17/16 11:32 PM (7 years, 2 months ago)

If you plan on growing long term, it would be foolish not to learn agar.
While your jars are colonizing I would gather supplies and make some no pour pasty plates to prepare cultures for your future grows:thumbup:


--------------------
:amanita2: Substrate surface conditions / Monotub prep and care :sporedrop:

Look around you... Everything you see exists inside the mind.
Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

Our individual experiences are all part of the universe's experience of itself

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Offlinekadmsu
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Re: Two Syringes: Here's my plan w/ questions [Re: ComebackKid]
    #23937042 - 12/18/16 01:02 AM (7 years, 2 months ago)

Th3Issu3: Definitely doing it, just trying to minimize the fuck ups. Thanks for the encouragement though :smile:!

ComebackKid: Thanks for the input on that.  I guess there is a balance between using a larger inoculation volume and trying to get a faster colonization rate vs using a smaller volume and reducing the chances of introducing contaminants as r.lutece eluded to.  Also, definitely planning on learning agar, just was gonna go from clone first instead of spores.

Thanks all!

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Offlinetump
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Re: Two Syringes: Here's my plan w/ questions [Re: kadmsu]
    #23937058 - 12/18/16 01:16 AM (7 years, 2 months ago)

A good idea is to also plan 6 months a head. Plan out how much you need to grow for you needs. Then plan on avg Yeild. Make double the amount of agar plates youll need to knock up that many jars as needed. Therefore even if you fuck up 50% of the time you will still get what you need

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