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OfflineMycobro420
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Clean spawn
    #26828546 - 07/17/20 09:29 AM (3 years, 6 months ago)

I recently got into cloning on agar. I take flesh peice an out to agar.then transfer it to qt jars.agar always comes out looking great rizomorphic growth rarley aerial myc. Fully colinize dish in about 10 days jars recover from shake.then I out to monotubs some of the substrates fully colinize some dont. One sub got trick before first flush.so my question is if my agar shows no visible signs of contamination and recover from shake and fully colinize bags or jars how an I getting trick/week colinzation in tubs.i had a bunch of bins trick from understerlized bags so I think my growroom is infested with trick is that possible or do I need re evaluate my steril tek


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InvisibleRoger Clemency
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Re: Clean spawn [Re: Mycobro420]
    #26828555 - 07/17/20 09:36 AM (3 years, 6 months ago)

How many transfers away from the clone plate are you going before putting it to grain? You should go at least one extra xfer from the clone plate. I usually go 2 or 3.  You can have recovery with bacteria and mold, though it’s usually a little slower but you might not notice it.

I saw a little grey mold in a jar a while back and shook it anyway to watch. Had I not known beforehand I might’ve missed it because it did recover with the others but like 2 days late to 100% and I had to look hard to notice the little bits of gray throughout.


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OfflineMycobro420
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Re: Clean spawn [Re: Roger Clemency]
    #26828573 - 07/17/20 09:47 AM (3 years, 6 months ago)

I transfer from original plate like you said atleast one transfer. All plates look great.i don't know how to use the sight real well but if you go to my gallery you can see a pic of a plate I uploaded. so u can see how clean my agar is. In my amateur opinion it looks great.my g2g bags come back after a shake my qt jars all within 24-48 hours.i do plenty of research an studying all day every day I'm rather on this sight or working in projects but this one has me stumped


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OfflineMycobro420
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Re: Clean spawn [Re: Mycobro420]
    #26828583 - 07/17/20 09:52 AM (3 years, 6 months ago)

My g2g qt jars come back to fully colinize in about 14 days I use 1qt of spawn for 10 jars an they always fully colinize ever after a shake.after already g2g so where is the trick coming from.i work at my buddy's house I'm the one that researches an does everything he is my assistant. We had alot of trick from last cycle cause I improper stuffed my 23qt presto to the brim with grains in bags an understerlized 20 bags but they still fully colinized bags then took to bins some flushed some got trick before first flush my confused.can airborne trick be so bad it effects all my future cycles


Edited by Mycobro420 (07/17/20 09:53 AM)


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InvisibleRoger Clemency
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Re: Clean spawn [Re: Mycobro420]
    #26829579 - 07/17/20 06:40 PM (3 years, 6 months ago)

I've seen people saying on here they had such a high spore load that their grows were contaminating even with clean spawn. I don't know how bad it would have to be though because trich in particular is already everywhere.

What is your bulk sub and how are you preparing?


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OfflineMycobro420
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Re: Clean spawn [Re: Roger Clemency]
    #26830217 - 07/18/20 05:56 AM (3 years, 6 months ago)

Ya we had it pretty bad.and I have limited control over at the spot we do it at.i teach my buddy everything I know but he doesn't listen to well.
So we had alot of trick from understerlized bags an he let the trick mature in another room by the growroom so I think the matured and are sporulateing might be in air.but on the other hand I have really Ben stressing cleanliness to him so it's looking alot better


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OfflineMycobro420
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Re: Clean spawn [Re: Mycobro420]
    #26830222 - 07/18/20 06:02 AM (3 years, 6 months ago)

My bulk substrate is standard recipe one brick of coir 2 qts of verm 4.5qts water handful of gypsum. 5qts spawn.all spawn was inoculated with 1/4 to 1/2 of agar transferred and fully colinized.they recover from shake some jars will be done in 17-21 days some 30 days all fully colinize in qts.thwn I do g2g to bags or just qt jars an all my spawn fully colinizes. Out of 80 jars and most with g2g and 5 bags all fully colinized but one jar.im kinda stumped


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OfflineMycobro420
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Re: Clean spawn [Re: Mycobro420]
    #26830225 - 07/18/20 06:04 AM (3 years, 6 months ago)

One more thing I see growth from shook jars within 24-48 hours if they take longer than that they usually don't fully colinize


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InvisibleRoger Clemency
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Re: Clean spawn [Re: Mycobro420]
    #26830348 - 07/18/20 08:36 AM (3 years, 6 months ago)

That's a lot of water in that recipe. I know some TEKs do say to use 4qt but I use 2.75 qt of water for the same amount of coir/verm. I do mine a little on the drier side like in the SFF shoe box method and mist the top layer after making the tubs. You don't get any side pins really that way.

I would try a little drier recipe which will let the myc colonize the bulk a lot faster and scale down your grows for a while. Like start new spores, transfer twice or so until you're getting clean plates with no contamination riding along or in satellites on the plate itself, then drop half plates into qt jars (cut up in to several wedges if you like) and then do some shoe boxes with a 1:1.5 ratio. You'll get fast colonization, about 10 days, pins in 10 more.

Instead of trying to expand your culture with g2g and dealing with bags and all that I'd try some smaller more simple grows for a little bit.


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InvisibleLadysKnight
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Re: Clean spawn [Re: Roger Clemency]
    #26830356 - 07/18/20 08:45 AM (3 years, 6 months ago)



Looks good backlit, what does the surface look like?


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Offlinesouthbounpachyderm
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Re: Clean spawn [Re: Mycobro420]
    #26830363 - 07/18/20 08:51 AM (3 years, 6 months ago)

Quote:

Mycobro420 said:
One more thing I see growth from shook jars within 24-48 hours if they take longer than that they usually don't fully colinize




I personally see rebound on most shaken stuff within 16 hours-24 hours. I don't trust anything that takes over that amount of time to fuzz back up. You can absolutely still have a bacterial infection and it will rebound. Only the really terrible bacterial presence wont rebound.

I completely agree with roger. stop trying to expand your cultures thru g2g and using bags. g2g is a semi advanced method and should really only be utilized when you are certain of what clean spawn and inoculate is. Bags are a whole other beast and they took me about a year to fully dial in after I had already been growing for a year by that point. Absolutely do not use LC or LI either. It seems like you're trying to take the easy route without putting in the time and effort.

First flush trich means you are having a catastrophic failure somewhere along the line. Do you have a flowhood? If you don't and you are using a SAB I would absolutely stop doing bags until you have one. That could be where your issue lies. Airborne trich can be bad and its possible thats the issue.

Just judging from your methods you're probably expanding lightly bacterial culture g2g and the myc is so weak upon spawning your tubs that the trich is able to overcome its defense mechanisms because of the bacterial presence.

G2G expansion is a force multiplier for semi bacterial spawn. You are introducing damaged myc to a completely new food source. Any bacterial presence gets the jump on that mycelium and can easily go out of control.

Do you have some pics of your bags, jars, and plates to share so we could potentially chime in on whether we think its clean?


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Offlinesouthbounpachyderm
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Re: Clean spawn [Re: LadysKnight]
    #26830365 - 07/18/20 08:53 AM (3 years, 6 months ago)

Quote:

LadysKnight said:


Looks good backlit, what does the surface look like?




There are a few spots in every transfer that indicate bacterial presence on this plate in my opinion.


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OfflineMycobro420
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Re: Clean spawn [Re: southbounpachyderm]
    #26830701 - 07/18/20 12:35 PM (3 years, 6 months ago)

Ok let me start by thanking everyone for chiming in.i have Ben doing this for about a year now. I have done a bunch of research into this hobbie an have plenty of free time to do myco work/research.ok to start I follow that recipe for my substrate because it's the same frank horrigan uses my sub with that recipe comes out the feild capacity. A few drips of water with a hard squeeze if I go under that recipe I can't get the few drops I'm looking for.i only use 66qt monotubs for all my subs.the depth comes to about 4 inch's.never really tryed less maby it would be beneficial I shall try that.im fairly new with cloning to agar.i have ran alot of spore syringes to qt then to mono with great success.yeild average are 112 for first flush 60 for second.very minimal contams with spore syringes.niw with the agar looking bacterial I understand why you would see it like that with the background light an no vids of the top of the dish.iv seen bacteria on dish's it has a milky look that is completely diff from clean looking myc.i wish I had a top picture but since that dish has Ben put to grain.with the jars taking 24 hrs to recover I don't live at the place where I do this so I can't monitor it 24/7 but most do show growth in 24hr  unable to look at then before a time frame under 24hrs. Wen I do more bags an jars I'll upload them an I have had ok success with agar my last cycle beat fruiter was just over 6oz dry first flush and 4 1/2 of second so I kinda know what I'm doing but looking to prefect my Tek so I don't have to worrie about where I went wrong.i also do own a flowhood I have had it for 9 months an do all steril work infront of it


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Offlinesouthbounpachyderm
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Re: Clean spawn [Re: Mycobro420]
    #26830784 - 07/18/20 01:27 PM (3 years, 6 months ago)

Not all bacteria looks the same and lots can be hidden when the myc grows over it.


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OfflineJus
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Re: Clean spawn [Re: southbounpachyderm]
    #26830852 - 07/18/20 02:05 PM (3 years, 6 months ago)

Quote:

southbounpachyderm said:
Quote:

LadysKnight said:


Looks good backlit, what does the surface look like?




There are a few spots in every transfer that indicate bacterial presence on this plate in my opinion.




I'm new to agar myself and would love to hear what you're looking for/where?

My untrained eye: The empty disordered patches in the 3 o'clock position of the top right transfer, and the similar empty patches dotted around the top of the bottom sample, but what about the top left?


Edited by Jus (07/18/20 02:06 PM)


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Offlinesouthbounpachyderm
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Re: Clean spawn [Re: Jus]
    #26830866 - 07/18/20 02:12 PM (3 years, 6 months ago)

Quote:

Jus said:


I'm new to agar myself and would love to hear what you're looking for/where?

My untrained eye: The empty disordered patches in the 3 o'clock position of the top right transfer, and the similar empty patches dotted around the top of the bottom sample, but what about the top left?



3-6pm all is iffy GOOD EYE tho. You've got this shit down


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OfflineMycobro420
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Re: Clean spawn [Re: southbounpachyderm]
    #26831019 - 07/18/20 03:32 PM (3 years, 6 months ago)

I have another plate actually I can post I put it in the fridge in insulated lunch box for storage an going to transfer to new plates soon.ill post it a bit later I don't know how to upload it onto threads but I can to my gallery.so you can see the top of plate.an ya I know bacteria can hid under but I'm on this site all the time looking at good treads from TCs and I don't see any visible problems I'll upload the other an let me know what you think.


Edited by Mycobro420 (07/18/20 08:59 PM)


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OfflineMycobro420
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Re: Clean spawn [Re: Mycobro420]
    #26832261 - 07/19/20 10:08 AM (3 years, 6 months ago)

Ok yesterday I checked agar plate I only have one at my house cause I do my myco work else where.the dish I have here was not properly stored.the insulated lunch box at other place.so I wraped this one in a towl an out it in fridge just to take it out yesterday an see a bunch of condensation inside dish.(probably unusable now)I can still post pic really hard to see.but u did upload to my gallery my original dish I took transfers from if you guys could take a look would be much appreciated


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OfflineMycobro420
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Re: Clean spawn [Re: southbounpachyderm]
    #26832291 - 07/19/20 10:31 AM (3 years, 6 months ago)

Also why would you say that lc or li is bad.i recently did my first ll attempt I used 150mi of sterlized water droped whole Petri dish into it.shook up didn't break up to well cause I use stiffer mea. I inculated 4qts if rye in a spwan bag. The bag fully colinized in 14 days. I spawned it to a monotub.the tub fully colinized in 9 days first flush coming in now.i got busy doing other myco work an didn't case the tub. (Species is pe) but it blobing away currently lol.please explain why u think lc an LL is cutting corners.


Edited by Mycobro420 (07/19/20 10:33 AM)


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InvisibleRoger Clemency
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Re: Clean spawn [Re: Mycobro420]
    #26833086 - 07/19/20 07:19 PM (3 years, 6 months ago)

It's not a bad method or cutting corners but if you're having contamination problems it's usually best not to mess with it because you're putting all your eggs in one basket while you have a hole in your game somewhere.


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OfflineMycobro420
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Re: Clean spawn [Re: Roger Clemency]
    #26833243 - 07/19/20 09:04 PM (3 years, 6 months ago)

I think I'm doing to many qt jars and my an alot of agar to put in each one so my cleanliness slips doing so much. I'm going to upgrade my pc as well.iv had mixed success with bags and agar.this is first time doing so many qts with agar. most look good too soon to tell I'll update wen more jars and bins are fully colinized


Edited by Mycobro420 (07/19/20 09:05 PM)


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OfflineMycobro420
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Re: Clean spawn [Re: Mycobro420]
    #26833251 - 07/19/20 09:11 PM (3 years, 6 months ago)

Why I was thinking about lc is I would pick my best plate and cut wedge an place it in lc to insure it's clean in front of flow hood. Then use it to inoculate bags I figure less contam risk because I only have to grow the lc out from little peice of myc instead of having to do alot of dishes with alot more transfer peices and diff agar drops.and instead of using lc syringe inoculation dump it into each bag


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InvisibleRoger Clemency
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Re: Clean spawn [Re: Mycobro420]
    #26833771 - 07/20/20 07:48 AM (3 years, 6 months ago)

I personally use LI for all my jars now. I love it. I do have an LC ready to go as well but with LC you have to worry about keeping it clean after you inoculate your spawn, until you’re down to the last of the LC.

With LI you just need a clean plate and a jar of sterilized water and you make it up right before inoculation, use it up and that’s that.


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OfflineMycobro420
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Re: Clean spawn [Re: Roger Clemency]
    #26833826 - 07/20/20 08:40 AM (3 years, 6 months ago)

I used LL like I said earlier for one bag with 4qts in it fully colinized in two weeks I shook it about 7 days in. It colinized bin in 9 days and is fruiting now.but I'm convinced that I'm doin to much at once being so new to agar. IV done 100s of qts of rye with spore syring with almost no failer.then did small run with agar went well now I'm trying to do alot more lolol.so first bigger grow with agar I did around 20 bags all went well colinzation wise.some bags had dry grain on top each bag got 5 hydrated qts an I g2g with whole qt wen I put them to monotubs around half tricked after or before first flush but I figured the trick came from understerlized grain cause I only own a 23qt preso PC and tryed to do 40qts of grain in one pc with no spacers. It was totally filled.still got good yeild from ones that made it.so this time I did alot more agar an qts cause I figured my contamination was from understerlized grain cause my agar looks ok check then new pic I uploaded it has rizomorphic grow on first dish.so I did this cycle with half qts and bags with only 4qts each bags did alot better and qts are doing ok to. But to soon to tell.


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OfflineMycobro420
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Re: Clean spawn [Re: Mycobro420]
    #26833833 - 07/20/20 08:47 AM (3 years, 6 months ago)

And with the lc my idea is to use it all at once.so with LL is same amount of precaution as lc in my eyes.but with LL you know exactly the amount of myc going into water.with lc you re grow the myc out so in comparison 150mi of grown out lc would have much more active growing myc.so it's like having 5 dishes in little jar an I would dump the jar from bag to bag.so instead of having agar dish cut 4 ways to 4qts of grain that's 4 diff chances for something to go wrong wen I figure if have one grown out jar I can dump into 20 bags so I know all 20 bags came from the leading edge of a Petri dish.instad of using a bunch of wedges and g2g.it would also save time instead of waiting for qt jars to fully colinized to even be able to do g2gs


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