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algeezy
Stranger

Registered: 01/07/22
Posts: 58
Last seen: 2 years, 8 months
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Stunted growth on clone in Petri dish? Should I transfer?
#27772905 - 05/11/22 04:57 PM (2 years, 8 months ago) |
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About three months ago, I harvested some GT and Z, took the best specimens, and attempted to clone. Growth has been virtually non existent. The first month had very little growth. I came on here and asked for help but didn’t really get any. I initially kept my dishes in my closet at 65F - 75F. People on this site said to incubate them at 80F-85F so I went ahead and got an incubator. After a month of seeing even less growth in the incubator and not getting any actual help from anyone, I gave up and haven’t monitored progress for almost 2 months. The pic below is the current state of the seemingly surviving plates. I used mss on several plates and saw 0 grown on 10 plates. A couple of the cloned samples have like black mold around them. Seems like there’s slightly more growth than what I saw 2 months ago but it’s hard to tell since I can’t really remember what they looked like before, but it’s not that much different.



My question is, should I just restart the whole process? If so, what can I do different?
I use PDA instead of traditional agar. Should I switch?
Any help would be appreciated. I have several mss so planning on using PF tech again to clone anyways, but trying to salvage the samples I do have.
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hazyhorse
scoobin



Registered: 03/19/19
Posts: 3,822
Last seen: 2 months, 5 days
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Re: Stunted growth on clone in Petri dish? Should I transfer? [Re: algeezy]
#27772914 - 05/11/22 05:09 PM (2 years, 8 months ago) |
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you don’t need to incubate anything in this hobby, at least relating to cubes & most actives. idk who told you that but they were wrong. i’ve colonized plates from 59f-86f & never had any problems with any temps. those plates look unhealthy & 2 months is waaaaay too long without anything happening. clones should take off within a week ime.
PDA vs LME agar or whatever shouldn’t matter. how did you take the clones initially? if you take them improperly they can fail hardcore
i’d go back to spore & do agar to grain jars or straight up PF tek (or both). you’ll get mushrooms from the MS & should have plenty of candidates for cloning
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algeezy
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Registered: 01/07/22
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Last seen: 2 years, 8 months
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Re: Stunted growth on clone in Petri dish? Should I transfer? [Re: hazyhorse]
#27772953 - 05/11/22 05:48 PM (2 years, 8 months ago) |
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Thanks for the info! It was my first time cloning but I pretty much did what the guides here say. Basically take a sample, and take the tissue from inside the stalk and place it in agar. I use the same sterile practice as PF tek
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hazyhorse
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Registered: 03/19/19
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Last seen: 2 months, 5 days
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Re: Stunted growth on clone in Petri dish? Should I transfer? [Re: algeezy]
#27772956 - 05/11/22 05:52 PM (2 years, 8 months ago) |
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assuming you did it in an SAB/flow hood yeah that sounds pretty standard. you could try mixing your own agar, either PDA/LME/whatever tek looks good to you & see if different nutrient ratios give you different results. do you put spores to your agar? or was the cloning the first time you used it?
-------------------- you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================
 
🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿
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algeezy
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Registered: 01/07/22
Posts: 58
Last seen: 2 years, 8 months
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Re: Stunted growth on clone in Petri dish? Should I transfer? [Re: hazyhorse]
#27772957 - 05/11/22 05:54 PM (2 years, 8 months ago) |
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So is that a no on trying to transfer?
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algeezy
Stranger

Registered: 01/07/22
Posts: 58
Last seen: 2 years, 8 months
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Re: Stunted growth on clone in Petri dish? Should I transfer? [Re: hazyhorse]
#27772958 - 05/11/22 05:54 PM (2 years, 8 months ago) |
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I used mss on some and used the ref for cloning
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hazyhorse
scoobin



Registered: 03/19/19
Posts: 3,822
Last seen: 2 months, 5 days
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Re: Stunted growth on clone in Petri dish? Should I transfer? [Re: algeezy]
#27772961 - 05/11/22 05:57 PM (2 years, 8 months ago) |
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i mean you could try to transfer, but it doesn’t really look like there’s much of a leading edge of mycelium to transfer from. you could try cutting out the thickets white spot you can find & put it to a new plate but idk that i’d expect much if they grew for 2 months & that’s all you have. you could have started over from spores & had a harvest within that time
did the multi spore plates grow myc out normally?
edit: just read that you said 0/10 agar plates grew from the syringes. i would maybe ditch the premade mix & try a recipe on here that has been proven to specifically work for mycology
-------------------- you're not the first to set foot here, just another
===================================
i love glass petris & you can too!!
posts i constantly refer back to
new to mushroom cultivation?? read this!!
===================================
 
🅃 🄴 🄰 🄼 🄲 🄻 🄸 🄽 🄶 🅆 🅁 🄰 🄿
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WyoMX

Registered: 07/06/15
Posts: 2,351
Loc: PNW
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Re: Stunted growth on clone in Petri dish? Should I transfer? [Re: algeezy]
#27772962 - 05/11/22 05:58 PM (2 years, 8 months ago) |
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I never had that good of luck with the premixed agar. In theory it shouldn't make a difference but when I bought my own agar agar powder and nutrients ( I use light malt but plenty of people use potato flakes and are successful as well ) my plates just seemed to work so much better and actually grow what I was trying to grow which is mycelium. Also what did you wrap the plates with?
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