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InvisibleComebackKid
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Interactive Guide to Cleaning Cultures on AGAR * 5
    #24473673 - 07/10/17 07:55 PM (6 years, 8 months ago)

Cleaning Cultures with ComebackKid
An Interactive Guide to Cleaning Cultures on Agar for Beginners


Introduction
In this hobby AGAR is the first step to cultivating and the only way you can expect to be taken seriously as a cultivator.

It's important to clean cultures on agar before moving them to grain spawn so that bacteria and other contamination won't have a chance to dwindle your yields or sabotage your grow altogether.

Agar gives you the ability to physically remove clean healthy mycelium away from bacteria and mold by growing everything out on a 2D surface and transfering chunks of mycelium to new petri dishes.

The good news is that it's incredibly easy and relitively cheap to get started, so if you have been putting off learning how to work with agar now is the time to get started.

I'm hoping to do this as a grow along where beginners can ask any questions that they may have during any part of the process. Everything from finding supplies to identifying mycelium.
I also encourage experienced cultivators to participate as well so we can help as a community
This guide will focus on

1. Supplies
2. Preparing your agar
3. Inoculating your agar
4. Cleaning cultures on agar
5. Inoculating grains with agar



Getting Started
Here are a few links for getting started with agar.

Pastywhyte's easy agar TEK This is a guide that will teach you how to prepare agar "petri dishes" from common household items.
Everything in this TEK can be found at your local Walmart and health food store.
I will personally be using "Pasty Plates" myself, but you will find that many people also prefer to use actual petri dishes. Both work just as well.

BOD's Simple SAB TEK This guide will teach you how to prepare and use a SAB (still air box)
This is the box where you will do all your sterile work (transfers, inoculations etc)



Supplies
A few things you will need to get started

-Still Air Box (at least 55qt clear tub)
-4" hole saw or empty coffee can
-Butane jet lighter or torch
-Scalpel or craft knife
-Pressure Cooker
-4oz pp5 tupperware (glad mini rounds)
-Micropore tape
-1/4" drill bit or hot nail
-Paper towels
-Aluminum Foil
-Agar agar powder
-Potato flakes
-Sugar



Participants
Here is where I will post links to the posts in the thread where participants will update their progres.

if you would like to sign up for the grow along please post "My grow log" and a little bit about your current understanding of agar in the comments section.
That post will be your grow log and you will edit your original post each time you have made some sort of progress. I will link to your post in the OP as soon as I can.
The idea is to catalog your progress to help other people who wish to learn agar so I encourage everyone to document as much as they possibly can.


Bodhisatta
Buddha19er
Capmuncher
Cautionarytale
Comebackkid
Dactylium
Enlightenment
Hamington Loafburg
[url=/forums/showflat.php/Number/24506612#24506612]Jeebo[/url
Kyshroomer19xx]
Leafygreens
Mr.Mushie402
Nesmaster01
PaddyWhite
SalviaGod
Senseit
Steevo
Streamer
Theoldclarysage
Wicked Burn
Willhelm

Edited by ComebackKid (08/14/17 06:29 PM)

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InvisibleComebackKid
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: ComebackKid] * 1
    #24490278 - 07/17/17 08:29 PM (6 years, 8 months ago)

ComebackKid's Grow Log


JULY 18
I've got my pasty plates made up using Munche's 123 PDA Recipe
This recipe is a bit more nutritious than Pastywhyte's recipe and the agar is softer which helps when germinating spores.


I'm going to be taking spores from a print  and putting them to agar with a cotton swab.

To start, I sterilized some cotton swabs in a mason jar and PC for 20 minutes.


In my still air box I picked up some spores from the print by lightly smudging it with the tip of the cotton swab.


With one hand, I cracked open and propped the lid of my plate, while the other swiftly put the spores to agar by swiping an X on the surface of the agar with the tip of the cotton swab.


Then labeled my plate with a permanent marker and placed on a shelf to wait for signs of growth.




JULY 21
I noticed some very faint growth in my plate. There is clear mycelium growing away from the X that I swabbed into the agar.


I outlined the leading edge in red so you can see better what I'm talking about here.
This is very early stage. Eventually the mycelium will thicken up and be white with the clear leading edge moving outward.




JULY 24
The mycelium had thickened up and you can clearly make out the white mycelium and clear leading edge.


Traditionally you would make transfers from the white leading edge between the red and yellow outlines.
I personally like to transfer from the clear leading edge between the red and green outlines.


----

Here is how I make my transfers.
I flame sterilized my scalpel blade with a butane jet lighter until it was glowing red.
Then, in a still air box, I sliced a small square into the agar where I planned to take my transfer from.


Then I scoop out the wedge by putting pressure on the side of the wedge with the tip of my scalpel.


Then I cradle it on the tip of my scalpel while I fetch a clean plate to transfer to.


While the wedge is being cradled I like to keep my scalpel hand steady and bring the new plate to the wedge. This way I'm not dragging my wedge through the air and possibly picking up contaminations on the way.
I place the wedge in the center of a new dish by dragging the tip of the scalpel through the new agar leaving the wedge behind.


--------------------
: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

Edited by ComebackKid (07/24/17 07:25 PM)

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OfflineBuddha19er
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: ComebackKid] * 1
    #24493555 - 07/19/17 06:37 AM (6 years, 8 months ago)

Skip's Grow Log

On this the 23rd day of July in the year of our Lord 2017, I have put Golden Teacher spores to agar!
I got all of my supplies together, and have been reading and watching videos during any free time I've had this week.
I used Munche's 123 PDA Recipe
https://www.shroomery.org/forums/showflat.php/Number/23188000/fpart/all/vc/1
And it seemed to go well.
I inoculated them with a GT syringe I had.
I put 2-3 drops in each one.
I did everything in my SAB, and used normal sterile techniques. My camera is messed upon my phone, so I will post pics tomorrow when my new phone gets here.

A little update...

These are some of the growths I've got going on. Don't know if they're good or bad yet. I used a spore syringe (from a reputable vendor of course). There are white spots appearing in the other plates as well.
Can I open the lid to get a better look and pics?
More pics. This is 5 days since inoculation ...


It's been a few days, and out looks like I got some good growth, and... a contaminate! I was kinda happy to see it, truth be told. Because I recognized it right right away from all the logs on here. Y'all have fun and let me know if there's something  you're seeing in these pics. Thanks

Been awhile, but that's life for ya. I'm on my second run of plates. I did 8 plates of a PE6 syringe and got contams in 6 so far. I have them separated and am going to try and clean them up tomorrow. Also, have 6 plates of SyZyGy that I got a print of yesterday and put to agar.
Hope everybody's plates are coming out clean and thick.
Oh by the way, I just spawned my first 5 quarts of oats that were Inoclated with my first agar plates Sunday. GT syringe>PDA>PDA> tiger dropped to Oats>Coir/Verm sub.

Edited by Buddha19er (08/15/17 05:03 PM)

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InvisiblebodhisattaMDiscordReddit
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: Buddha19er] * 1
    #24493776 - 07/19/17 08:50 AM (6 years, 8 months ago)

Some helpful agar videos

Grow Log: Interactive Guide to Cleaning Cultures on AGAR


8-26-17 O-GBWA (oat grain boil water agar) was used undiluted from the grain water obtained from using my oat tek. 20g/L of agar.

streaked some Matias Romero to agar

4 days later it's ready for a transfer


Put PESA clone to agar, and tried to be as un-clean about it as possible moving the plate around in the SAB by the arm holes. And while ripping the fruit open using my bare hands. I wanted a not clean plate for the purpose of this thread. It worked and I got a bunch of satellite mold colonies. And a few bacterial colonies in the middle.


here's a closeup the green circle is where the transfer is from. You can see the few bacterial colonies and the mold that's really close to the cubensis mycelium.

Here's the result of the 2nd plate



transferring another time just to play it safe.

9-15-17 here's the pesa on the 3rd transfer


here's some PE6 spores germinating on agar. I didn't do a streak for this one


and first generation prints of wild texas on their 2nd transfer going to their 3rd



9-15-17

3rd transfer, on wort agar at about 4% sugar content




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OfflineSteevo
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: bodhisatta]
    #24493811 - 07/19/17 09:10 AM (6 years, 8 months ago)

Steevo's Grow Log

Great idea CBK. Let's do this  :rockon:

I'm dumping these here for now but I'll be back in a while to follow up. I transfered a bunch of spores to PDA as per Pasty's recipe on 7/7 and 7/9, went away and found these when I got back home just now. They're in mason jars which is making it tough af to get a good pic of. I was using mini rounds before but switched to these for a slightly larger area to work off of. I'm definitely going to look into getting a sleeve of petri's asap

These top 2 are the tip I cut off of an APE swab



This was streaked with the swab before cutting and looks bacterial

  These are ATL7 but don't seem right to me. Looks like black mold



  These next few are from some Morrel spores sent to me from a member that I put to pda to see what would happen









I'll document shit a little more step by step in the next few days when I get settled and start a proper log, I just thought I'd dump some shit here for fun
-------------------------------------------------------------------------------------------------------------------------------------

7/23- So I went ahead and ditched anything suspect and just made up a bunch of no pour plates a la Pastywhyte. I used 6tbsp of agar, 1.5c of water and 4tsp of potato flakes. Added a few drops of green food coloring and voila!



So this is how I pc my shit. Whether mini rounds or mason jars I do it the same way every time

  I put a layer of jar rings on the bottom covered with a piece of foil that I slash holes in with scissors

  Then I put enough water in to come up to the 3qt mark which ends up being perfect for me. I bring the pc up to a boil without a lid on, put the lid on and vent it for 10min, then put the weight on and run it until it rocks at 15psi. At this point I can turn the burner down or just run it full tilt for 45min with no worries of it running dry. The steam does all the work
-------------------------------------------------------------------------------------------------------------------------------------

7/24-Went ahead and streaked 4 plates with a PE swab that a member was so generous to send me :thumbup:
May end up scraping and cutting off a small piece of the tip and putting that on a plate to see what comes up
So hard to get a good pic through these mini rounds



    I clipped the tip off of an APE swab and let it grow out a little too long but I took a few samples tonight anyway :shrug:

I honestly don't even remember doing this but its pretty bacterial already so its going in the bin

 
-------------------------------------------------------------------------------------------------------------------------------------

7/28- The samples I tried to transfer turned bacterial on me so they're going in the trash. I inoculated a few more plates today using a Mazapatek print sent to me by an awesome dude and did some more PE this time having scraped the tip with a scalpel and streaking the plates with it. Its really tough to get a good pic of these plates which is making me lean towards buying a couple of sleeves of petri dishes



I have a few of these Pesa plates that are starting to look promising so I'll keep my fingers crossed for them

Going to do up another batch of plates tonight. Planning on transfering a few of those stones from the morel plates and am going to also look into another recipe for stone producers. I have this ATL#7 print that I haven't been able to get to germinate for the life of me that's quickly becoming a pain in my ass lol
-------------------------------------------------------------------------------------------------------------------------------------

7/29- Some pics I took yesterday



Although I let it go a little too long I still transfered some samples from this plate that looked good as of last night
-------------------------------------------------------------------------------------------------------------------------------------

8/8- Have a lot going on and am waiting for a few free minutes to do a bunch of transfers :smirk:
------------------------------------------------------------------------------------------------------------------------------------

8/11- Made up a bunch of no pour plates yesterday with new containers I picked up from Big Lots that I saw a member post pics of. Doing a bunch of transfers today and got a couple of pics of some nice looking plates I'm taking from

PE    PE  B+ Clone
Pesa  Amazon    Tamp    Tamp

Going in for a rescue mission on this tamp plate

  These guys got overrun 
------------------------------------------------------------------------


8/14- A bit more growth going on after doing a few transfers. Top 2 on the right have not had a transfer done yet. Pc'ing plates now



------------------------------------------------------------------------


8/31- Alright so I haven't updated anything in about 2 weeks so here goes

All transfers are between 3rd and 5th at this point with a lot of the plates looking like these 2 here


I transferred wedges of good looking tampanensis growth on 8/24 into wbs jars and have my fingers crossed for truffles. Those 3 jars look something like this


I took a clean plate of PE and turned it into a Li and split it between 3 jars on 8/24. All jars look like this and will be left to consolidate for another week


I took a good looking oat jar of PE and used it to noc up another 6 jars of wbs on 8/29 and they look like this


I'll be going in to rescue this B+ culture a little later today along with starting some more varieties. I'm thinking ATL#7, LGT and Wild TX :rockon:

-------------------------------------------------------------------------------------------------------------------------------------

9/4- Found these 2 guys just chillin together at the back on the shelf
Tampanensis and PE
-------------------------------------------------------------------------------------------------------------------------------------

9/11- A bunch of cultures going but these ones stood out for one reason or another


Yet another little pinner
Tamp stones

Edited by Steevo (09/14/17 11:35 AM)

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OfflineBuddha19er
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) *DELETED* [Re: ComebackKid]
    #24493878 - 07/19/17 09:53 AM (6 years, 8 months ago)

Buddha19er's grow log

ComebackKid said:
I linked your grow log to your first post instead bro :thumbup:
Just make sure you edit that one when you update


--------------------

"The seeds that were silent all burst into bloom, and decay
And night comes so quiet, it's close on the heels of the day..."

Edited by ComebackKid (07/19/17 04:24 PM)

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OfflineJeebo
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: Buddha19er]
    #24493975 - 07/19/17 10:52 AM (6 years, 8 months ago)

Sign me up, bought my first spore syringe on monday and already have 3 agar jars going till my petri's arrive. Cheers for the help!

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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: Buddha19er]
    #24494135 - 07/19/17 12:39 PM (6 years, 8 months ago)

Perfect timing ComebackKid! Been having a little trouble with 2yo syringes.

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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: streamer]
    #24494363 - 07/19/17 03:08 PM (6 years, 8 months ago)

Capmunchers Cube Agar Adventure:

Wild Subsecotiodes Cleanup

18-7-17
So on a little walk I found a very strong growing  Subsecotiodes patch. I decided to take a few small pins see how hard it would be to clean up on agar.



This transfer was done with no real care to being super clean. The pins were washed with tap water, dried on a tissue and then dropped onto the agar in a SAB. Yes I am sure i could have dipped them in iodised or belched water but it was late, I was not planning on doing any wild cloning that day and I am lazy....

19-7-17
I can see lots of contams around the pins but sure it's mycelium that is running away.



20-7-17
This is a photo taken just under 24hours from taking the transfer from the leading edge of the wild pins plate. The plate I took the transfer from STUNK something evil but looks like I didn't bring to many nasties over with the samples.



To me the growth in the first plate looks super fast and clean...finger crossed! At the same time as taking the pin clone I tried some inner tissue samples from some good looking caps... Seems all i got was bacteria....

22-7-17
This thing is rocketing off so got some more transfers done.



8-8-17



14-8-17


made more transfers from the dirty plates but they still look slimy... having to have moved my setup it seems the drip fell onto the plates and sloshed things around. My hot pour attempts we not successful so gave up on this side project for now. I have a few buts that grew onto some cardboard that I might try again when the out door conditions will be right again.

--------------------------------------------------------------------------

Fun with Cubes

4-7-17
Made MS swipe plates of GT, AA+ and B+ using prints given to me by the kind members of this forum. Used an inoculation loop in a SAB on premade agar petri dishes to  take a zig zag swipe on each plate. I also did 2-3 plates of each print to make sure I had options should I have plates taken out by the nasties.

I made my own inoculation loop form a scaple handle using a bent bit of SS wire from the fishing shops. I used a Paper clip for a while but found it would hold LOTS more heat compaired to thinner SS wire. Vaping stores are an other option for cheap SS wire too.

I wrap each plate in two rounds of a cut down Gladwrap Brand plastic food film. Don't skimp here as the branded stuff is a touch thicker and sticks much better. To cut it down I get a BRAND NEW box cutter blade and put it up on a packer to hold it level and stable (tape it down to the packer if you want) Then I stand the Glad tube next to it and turn it so the blade cuts a grove, go slow and many times around and it will cut super clean and constantly. I feel this gives me a cleaner edge to the tape with less taggy bits.

11-7-17
Lucy - (Leucistic) Golden Teacher  |  AA+  |  B+


Decent growth on the Lucy (GT) and AA+ but the B+ is super slow. lost a plate to something green and evil. 1 bad from 7 is not to bad in my books :smile:

16-7-17
12 days in and getting bigger but still feel that the growth could be better, not sure if it could be my premade agar.... or maybe I'm being to rushed :blush:

Lucy


AA+


B+


21-7-17
A few photos at my first attempt to pour agar....



Thigs that worked - Agar media bottle, worth every penny as pours MUCH nicer than a beaker/flask. Need to order more. My method was clean as my control plates are still looking contam free.
Things that failed - the way I poured, moving the plates across and stacking in a pile beside was slow, poured while the agar was a little too hot and did it in a cold room. Ended up with wet drippy uneven plates....

25-7-17
Took transfers and then had to move house ... what a pain in the ass... Also suffering from a lot of condensation issues that looks like it creates wet plates. These photos are taken about 10 days after the transfer.



29-7-17
LGT latest transfers:


LGT older transfers:


Evil drips  |  B+


2-8-17
Got my shit together and poured some fresh plates. I made two batches to see what works best for me.



Red: half grain boil water (with a touch of gypsum) + half tap water + 0.5% LME and 2% agar

Green: all tap water 2% LME + 2% agar

This time I took the effort to filter the heated/dissolved agar solution as it was going into my media bottles before sterilization. Worth the effort as my plates are far more transparent this time with the GBW having some very small flecks and the LME being dead clear.

The first batch was poured at 49c but was just starting to gel on the last plate. The green I started pouring at 52c and found I was either getting faster or the extra temp gave me the time I needed. I also poured this stack from the bottom up, lifting the whole stack of emptys as I went, a bit more risky but much faster.

I hope to let these sit for a day or so and hope that the last few hazy dishes clear up. I tired a hot cup on the really bad ones but it seemed to just drop the water onto the top of the agar to make a wet looking surface.... Sill should have poured in a warmer room too... Next time...

8-8-17

Latest photos of my LGT plates.

14-8-17

Time to move cultures off plates that have POOLS of water .... I got to get this dripping situation under control.


Also time to take some transfers from a wild P.Subaeruginosa print.

15-9-17

Took some clones from the best fruit from my first few trays as well as a MS swipe fresh from the cap. No I will never keep a mold farm like that again....


17-9-17

The above AA+ clones a few days on:

20-9-17

First transfers away from the above plates growing out. Glad I went with lots of samples as a few are still a little bacterial.




Edited by capmuncher (09/20/17 05:35 PM)

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InvisibleComebackKid
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: capmuncher]
    #24494672 - 07/19/17 05:35 PM (6 years, 8 months ago)

Hey guys glad to have you on board
Just want to let everyone know that you can get started with your grow logs as soon as possible and to remember to edit your original posts with your progress.
I have linked to everyones posts who have signed up so far

buddha19er
How did finding your materials go?

Jeebo
Nice to see a new poster here! Have you had a chance to put the spores to agar in a SAB yet?

Streamer
What have you tried so far?

capmuncher
Wild samples can be a bacterial nightmare. It will be great to have em documented here thanks for joing!!


--------------------
: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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OfflineWicked Burn
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: ComebackKid]
    #24494771 - 07/19/17 06:13 PM (6 years, 8 months ago)

Wicked Burn's Log

I'm in.  Badass thread CBK.  :thumbup:

I've had success with germinating spores on agar from spore syringes.  This is my first experience making and using swabs.

I made PE swabs in my SAB with these individually wrapped sterile polyester tipped swabs.



With no dickin around I immediately put them to 12 pasty plates on July 13th.  I used the grain water recipe from Munch's Grain Water Agar thread.  Freshest spores I've ever used.  I just dabbed the center of the plate with the swab.  I probably should have streaked it.  A couple plates germinated and had visible growth in just 2 days.  Most of the plates had visible growth in 3 days and by day 4 all had visible growth.  Much faster than I've ever seen using spore syringes from vendors.



Can't see it in the picture above, but every one of those plates has a nice fluffy spot of myc in the middle.  But all of them are also surrounded by bacteria.  It seems like the bacteria is keeping the myc from growing outward any further.

Tomorrow I'll be trying to transfer half of these to new plates to try to clean up, and the other half I'll be trying a hot pour.  Anyone have any helpful tips or links to threads on the hot pour?  It'll be my first time attempting it.


Update: 7-20-2017

 

SAB, Alcohol dispenser, inoculation loop, butane torch, face mask and my favorite mits.



I used a soy sauce bottle to perform the hot pour.  I poured when my temp gun read 117 degrees F.



These are the completed hot pours.  Excited to see what happens.

     

Overhead shots of the plates I took transfers from.  I use an inoculating loop to take transfers.  I find I can be much more accurate and take much smaller pieces with the loop than with a scalpel.

Not sure where all the bacteria is.  I may have misinterpreted water streaks as bacteria or maybe it grew all the way to the edges making it even more difficult to differentiate...  Or maybe pasty plates are just tough to see real well and my eyes were playing tricks on me.  Or maybe I'm just loosing my fuckin mind.  :shrug:  Let me know if any of you see anything in the pics that I don't.

Here's the receiving plates after the transfers.



Update: 7-25-2017

So here is a picture of one of the hot pours.  I'm not really sure it's even mushroom myc.  Anyone else have any opinions?  Let's ignore the obvious contam near the side there.  The edges where I scraped todays transfers from looked better, but not much.



I'm starting to think I didn't germinate spores at all.  I guess I'll just keep transferring and see what happens.

One more thing, when I scraped the myc off the top it did do it's little shrinkage disappearing trick that i've seen cube myc do so many times.  Is that any kind of an indicator or does all myc do that?


Update: 7-29-2017

Here is what all the transfers I took from the hot pour look like today.

       

The growth is looking really nice and healthy.  The hot pour seems to have been extremely beneficial.

For comparison, here are a couple transfers that I took directly from the germination.  This is what they all look like today.

 

The NO hot pour transfers look the same as the original germinated plates (pictured earlier in the OP).  Just bizarre looking myc that doesn't grow outward, it just grows upward.

I'm suspecting there is some sort of invisible or difficult to see contam making the myc act odd.  The hot pour seems to have remedied this.  Awesome.

The hot pour will absolutely be something I'll be using regularly in the future.  I would recommend it to anyone with contamination problems.  Probably wouldn't work great on molds, but if you have bacteria or suspect a contam that you cannot see, go for the hot pour. 
:rockon:


Update: 8-5-2017

I took a whole bunch of transfers from these two plates today.

 

The growth is pretty faint, I've been using oat grain water agar.  I think I'm going to go to Munch's PDA on my next set of plates.  See if I can't get some more appealing looking growth.

I wasn't even 100 % certain I germinated and transfered cube myc until one of the hot pour plates started to knot up.  Check it out.

 


Update: 8-11-2017

That hot pour plate that was knotting up is pinning now.



The growth on all my plates from the last transfer is clean.  I've decided I don't care that it's not rhizomorphic.  I might as well keep as much genetic variance around as I can.  I'm going to make LI as soon as these plates are 100% colonized and then I'm going to knock up 9 masters and g2g from there.




Update: 8-14-2017

Here's what the pinning hot pour plate looks like today.



I looked at the bottom and found this orange splotch on the G and L of GLAD.



Could this be my suspected hidden contaminant that was causing the myc to act so odd before the hot pour fixed it?


Update: 8-21-2017

I made munch's blenderless LI and knocked up these 9 jars on 8-16.  The growth is clean and fairly quick.

   

I'm going to reiterate, if your myc looks odd and you suspect a hidden contam.... Hot pour it.  Works wonders.


Update: 8-25-2017

G2G'd these two masters into 10 quarts of oats. Once fully colonized I'll shake test em then spawn if clean.

 

Edited by Wicked Burn (08/25/17 04:32 PM)

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InvisibleComebackKid
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: Wicked Burn]
    #24494798 - 07/19/17 06:22 PM (6 years, 8 months ago)

Nice stuff Wicked Burn :rockon:
I have had success in the past by sandwiching the agar
and you could also try Tagar
Im currently trying this method out myself on some other cultures


Will you be able to grab ariel shots of your plates after you transfer and before you toss? just to get a better idea of whats going on in there?


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

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Consciousness, the awareness that is experiencing this mind,
is peering in from outside the universe.

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OfflineWicked Burn
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: ComebackKid]
    #24494838 - 07/19/17 06:38 PM (6 years, 8 months ago)

Yeah, I'll take overhead shots for sure.  These pasty plates have been used a fair amount so they're even less visible than they are normally.


--------------------
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Joe:  "Because you're a faggot."
                                     
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: ComebackKid]
    #24494858 - 07/19/17 06:53 PM (6 years, 8 months ago)

Had a thing to do for my kids, but have my list made up, and what stores to hit up. Should be home and starting by 7pm. Can't wait! Thanks again for offering this up to the community.  :bender:


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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: Buddha19er]
    #24494906 - 07/19/17 07:15 PM (6 years, 8 months ago)

I started watching the videos and and noticed they are making their plates from a print. I have syringes, but I don't have any prints. Will the syringes work, or do I need to get my paws on some prints? My wife had to throw the few I did have out, due to the fridge breaking down.


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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: Buddha19er]
    #24494968 - 07/19/17 07:56 PM (6 years, 8 months ago)

Syringes will work just fine for this :thumbup:
Just a couple drops is all you need


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

Look around you... Everything you see exists inside the mind.
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: ComebackKid]
    #24495052 - 07/19/17 08:51 PM (6 years, 8 months ago)

My grow log

Excited to be in on this. I just sterilized a few plates an hour ago.
My experience so far: started agar about a month ago. My real problem is identifying  bacteria in a culture. Either im an agar pro with very little contams or im missing something (and im assuming the later lmao)
My first bulk was a success, leaving me with a few nice gt prints. Tomorrow i will be putting these spores to 8 plates.

8/3
Wow, sorry guys I was swamped at work for the last tree weeks and never updated my log. On 7/21 i put spores to 8 plates. On 8/1 i made one transfer from 7 of the plates. One of them never germinated. One plate had a lot of rhizo growth so i took 2 from it. 8 transfers total.
Here are some pics of what my spores germinated into.

This one i took the transfer from about 5 o clock on the top colony.
1 o clock
11 o clock
11 o clock on the right colony. On the left i see black spots. Is this pin mold. Also the yellowing indicates bacteria correct?
this one i believe i scraped one of the ends of the larger strands at about 7:30. There are 2 thick longer strands and i scraped one from between them.
12 o clock.
9 o clock
The transfers were taken 2 days ago so the myc has grown a bit since then. I cant remember which one i took the second transfer from.:smbfacepalm: I scraped another rope of myc as previously described from one of those plates lol.


Took a break from prepping the next round of plates to type this up:stoned:
These prints im using were not taken in a clean manner. Can you guys point out some contams in the pictures in my update? Other than what i described my eye really just sees mycelium..... Lol.
How can i distinguish tomentose growth from contams? And what are the signs of bacteria? I know theyre loaded with it, these things smell SOUR, but how do i see it so i can take transfers farthest from it?

Edited by Mr.Mushie402 (08/05/17 11:39 AM)

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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: ComebackKid]
    #24495205 - 07/19/17 10:28 PM (6 years, 8 months ago)

Ham's log stardate supplemental.  These are the voyages of my experience with agar. 

I pour plates, and use flow hood.  I have also been known to employ no-pour methods using 8 ounce wide mouth squat, and regular wide mouth jars.  My favorite types of agar are grain boil off water agar, or malt extract agar.  I like to use 3 section plates for spore inoculations, and transfers, and single section plates mainly for clone tissue sampling.  I have a lot going on on agar right now.  From plates that are ready to be transferred from spores to plates that were just inoculated with spores, as well as, clone tissue culture that is ready for transfer. 
     

   


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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: hamloaf]
    #24495249 - 07/19/17 10:51 PM (6 years, 8 months ago)

SalviaGod's Grow Log

So ill start off by saying that im much more of a lurker than i am a poster, i think thats obvious based on my sign up date to post count ratio.
Im very new with agar, but in my short amount of time working with it iv made about 30+ transfers inside my SAB and have only ever had two plates come out contaminated.

Im going to cook some fresh agar up tonight and try to have some photos of my process up soon.

ALL POINTERS, TIPS, CRITICISMS, INSULTS( OR WHATEVER THE HELL YOU WANNA SAY ABOUT MY TECHNIQUE) ARE WELCOME 

Update 07-20-17


For my agar plates i use the Glad Mini Rounds that are used in Pastywhytes easy agar tek.
I melt a 1/4th inch hole in the lid and cover with two layers of micropore tape.


My agar recipe is a little wonky i suppose because i use a premix MEA, but i dont have any idea what its ratios of nutrients to agar are, but based off the growth of my previous plates, i think that its nutrients are too high, so i cut it with some grocery store agar.


For these plates i used 3 grams of the premix MEA and 2 grams of the store agar and 150ml of water.
I also add in two drops of green food coloring to help contams stand out on the plate.

This recipe will fill 8 mini rounds.

I like to fill the agar to the bottom of this squiggly curve that wraps around the plate, i use this as a guide line so i put about the same amount of agar in every plate.


(I usually let the agar cool in the plate before i cook it so i can test its consistency and firmness)

I wrap the tops in foil then PC at 15 psi for 45 minutes


After the PC i put them in my SAB


On these plates i swiped sterelized cotton swabs on prints and swiped an S shape onto the agar.
4 plates were inoculated with Pan. Tropicalis and the other 4 were inoculated with B+ spores from a print i took off my first grow ever back in 2013. Since i was a super noob back then the print is likely very dirty.


Update 07-25-17

After just 5 days the pan. Tropicalis are showing alot of growth that appears to be healthy.

Sorry for the picture quality, its hard to take photos with a phone with all the condensation on the walls.
No signs of other mold, or bacteria as far as i can tell. Im gonna cook up some fresh agar tonight and make some transfers. (Is one transfer per plate standard or do i need to make several? Im trying not to let this get out of hand by exponintially multiplying each plate and ending up with hundreds by the end of this.)
The B+ spores are showing no signs of growth yet, im assuming because they are old spores and probably need to be rehydrated.

Update 07-26-17

I decided to wait until today to do the transfers because i was tired yesterday.
Apparently that was a bad idea, as all my plates have grown more than i should have let them.
The S shape i originally swabed grew into a big blob, and one of the plates became covered in pin mold overnight.

Other than the one pin mold plate and the overgrowth, the transfers went smoothly.
I still took a transfer from the pin mold plate the best i could, just for practice i guess.
I dont mind throwing it away if its a lost cause, just figured id give it a try.


Edited by SalviaGod (07/26/17 07:16 PM)

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OfflineJeebo
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Re: Interactive Guide to Cleaning Cultures on AGAR (Sign Up Now!!) [Re: SalviaGod]
    #24495676 - 07/20/17 04:59 AM (6 years, 8 months ago)

Yeah although my sab is seriously small I started 3 agar jars with combobia, only messed 1 up by ingecting way too much spore solution, will get picture's up soon.

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