Home | Community | Message Board


This site includes paid links. Please support our sponsors.


Welcome to the Shroomery Message Board! You are experiencing a small sample of what the site has to offer. Please login or register to post messages and view our exclusive members-only content. You'll gain access to additional forums, file attachments, board customizations, encrypted private messages, and much more!

Shop: Mushroom-Hut Liquid Cultures   PhytoExtractum Buy Bali Kratom Powder   North Spore Injection Grain Bag   Left Coast Kratom Buy Kratom Extract   Original Sensible Seeds Bulk Cannabis Seeds

Jump to first unread post Pages: 1 | 2 | Next >  [ show all ]
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
My first grow, and I'm having some issues.
    #27041014 - 11/15/20 05:59 PM (3 years, 2 months ago)

Hi there, I need to ask for some help.

I've been reading everything I can on the forums for the last couple months but now that I'm actually starting I need another set of eye's to keep me on the right path. So here we go, I made a SAB, ordered MS syringes from a sponsor, purchased everything I could think of to complete the process, and repurposed a room in the house that I can keep Fairly sterile, and Control the Temperature. I decided to try MS to pre-sterilized bag (we're not going to talk about that one much) and also to do MS to agar. My first plates weren't gems to say the least but they're clean, or at least they were. I made a second round of plates with a little more care to resolve the issues I had with the first ones and they came out great. So, I thought I was doing pretty well until this morning when I looked at my plates and convinced myself they were most likely garbage. Everyone get some popcorn and prepare for a good laugh. this will be photographic evidence of my impatient noobery.

1.I have about 16 dishes like this, all of them have the same pattern developing

http://

2.
http://
3.
http://
4.
http://

That should give you an idea of what I'm looking at. for laughs, this next one is me not believing that anything was actually being transferred with the loop and decided to just put my (gloved)finger in the spores and go BOP!!! right in the agar. fuck it, right? I'm doing this with my daughter, and we're having a ball.

5.
http://

ok, so now that you've seen our plates I need some opinions on whether or not they are garbage or salvageable? as you can see they were inoculated on 11/12 and it's 11/15 now. I have 19 new plates made up (#20 was a sad yet hilarious loss to warpage after my daughter said just use the torch to heat it up to get rid of the condensation!) and 500 more on the way this week. My plan is to follow the wisdom I've learned here and make enough transfers to obtain a clean Petri which I'll use to A2G transfers.

So what do you all think about what you see? Garbage? wait it out?

I have more Syringes on hand, but I just cant think of what would cause that pattern on the plates. I know my plates are clean ( I have a control) and I followed Bod's sterile Tek video on making print transfers, I just substituted the spore print for a sterile petri with the spore solution in it. Hey, at this point I just want to say you guys make that shit look easy. working in a SAB and being conscious of every movement you make is no easy task.

My thought is that its probably a loss at this point. Daughter says wait it out. I just don't know enough to discern new Myc growth from contamination Or the Time Frame I should be looking at with making a Transfer to start clean up?

oh yeah, temp is constant 75F and 12/12 light cycle at 6500K

lots to unpack there I know, I'm really in no hurry, I'd rather get it right than get it fast at this point. I'm loving the science involved in the hobby and want to get learn from my mistakes.

Thanks to everyone that's put up so much time and effort to make this resource what it is!

FJ


Edited by Frozen Jedi (11/15/20 06:12 PM)


Extras: Filter Print Post Top
InvisibleRafiikii
 User Gallery


Registered: 11/17/10
Posts: 2,891
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi] * 1
    #27041021 - 11/15/20 06:05 PM (3 years, 2 months ago)

Your plates are contaminated with bacteria and I see no visible mycelial growth.

You are flaming your loop right?

I would recommend working on your sterility.

Such little spore is needed in this hobby.

We all make mistakes, learn from it and try again.

Good luck


--------------------
"You didn’t come into this world. You came out of it, like a wave from the ocean. You are no stranger here."



Edited by Rafiikii (11/15/20 06:10 PM)


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: Rafiikii]
    #27041042 - 11/15/20 06:15 PM (3 years, 2 months ago)

Yes, dipped in 70% ISO, Flamed red (outside SAB) scrapped in dish with spores (in SAB) then into the dish.


Extras: Filter Print Post Top
InvisibleRafiikii
 User Gallery


Registered: 11/17/10
Posts: 2,891
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041048 - 11/15/20 06:20 PM (3 years, 2 months ago)

Flame between each petri?


--------------------
"You didn’t come into this world. You came out of it, like a wave from the ocean. You are no stranger here."



Edited by Rafiikii (11/15/20 06:21 PM)


Extras: Filter Print Post Top
Offlinekanemush
Grumpy
Male User Gallery
Registered: 12/07/17
Posts: 1,502
Last seen: 9 months, 16 days
Re: My first grow, and I'm having some issues. [Re: Rafiikii] * 1
    #27041052 - 11/15/20 06:23 PM (3 years, 2 months ago)

did you keep the plates wrapped before and after inoculation?


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041065 - 11/15/20 06:34 PM (3 years, 2 months ago)

after inoculation i wrapped them. I'm confused on how the pattern of obvious contams starts towards the end and then follows the trail of the loop's path. also the rest of the plates seem are clean.


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: Rafiikii]
    #27041068 - 11/15/20 06:35 PM (3 years, 2 months ago)

Quote:

Rafiikii said:
Flame between each petri?




I did 2 plates per flame cycle


Extras: Filter Print Post Top
InvisibleRafiikii
 User Gallery


Registered: 11/17/10
Posts: 2,891
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041072 - 11/15/20 06:38 PM (3 years, 2 months ago)

flame every plate.


--------------------
"You didn’t come into this world. You came out of it, like a wave from the ocean. You are no stranger here."



Edited by Rafiikii (11/15/20 06:38 PM)


Extras: Filter Print Post Top
InvisibleRafiikii
 User Gallery


Registered: 11/17/10
Posts: 2,891
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi] * 1
    #27041074 - 11/15/20 06:40 PM (3 years, 2 months ago)

Quote:

Frozen Jedi said:
after inoculation i wrapped them. I'm confused on how the pattern of obvious contams starts towards the end and then follows the trail of the loop's path. also the rest of the plates seem are clean.




you took the bacteria along for a ride through out the whole streak

ISO your gloves/ consider some tyvek sleeves


--------------------
"You didn’t come into this world. You came out of it, like a wave from the ocean. You are no stranger here."



Edited by Rafiikii (11/15/20 06:42 PM)


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041077 - 11/15/20 06:43 PM (3 years, 2 months ago)

assuming (I believe correctly) that these are total losses, whats my best course of action to go from syringe to agar. There's lots of ideas about this and I haven't decided on the next plan.

One drop in the middle of a plate, wrap it and let it sit?

one drop in the middle then drag a loop through it? this seems like it has the most contam vectors but eliminates the blob of growth in the plate.

option 3- dealers choice?

I still have fresh syringes to use but won't have a clean sterile dish until this week sometime from 11/17-11/20


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041087 - 11/15/20 06:46 PM (3 years, 2 months ago)

flame every plate- check
Gloves and ISO wipe I did up to the elbows. I'll get some sleeves, do you reuse them? wipe them down? or one and done?


Extras: Filter Print Post Top
OfflineCocaineBuffet
Stranger
 User Gallery


Registered: 08/29/19
Posts: 3,488
Last seen: 10 hours, 35 minutes
Trusted Cultivator
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041092 - 11/15/20 06:50 PM (3 years, 2 months ago)

Personally I would go with the single drop and streak across the plate with the inoculation loop. Syringes just make the inoculation process convoluted. Most users here use prints/swabs.

Take things slow when working in the SAB. The more controlled and slow movements the less the air in the SAB gets stirred up.


Extras: Filter Print Post Top
InvisibleRafiikii
 User Gallery


Registered: 11/17/10
Posts: 2,891
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041093 - 11/15/20 06:51 PM (3 years, 2 months ago)

yeah definitely re usable, shit back in the day one would rock a whole tyvek suit. haha.

pretty cheap at home depot or lowes, in the paint section.


--------------------
"You didn’t come into this world. You came out of it, like a wave from the ocean. You are no stranger here."



Extras: Filter Print Post Top
OfflineCocaineBuffet
Stranger
 User Gallery


Registered: 08/29/19
Posts: 3,488
Last seen: 10 hours, 35 minutes
Trusted Cultivator
Re: My first grow, and I'm having some issues. [Re: Rafiikii] * 1
    #27041105 - 11/15/20 06:59 PM (3 years, 2 months ago)

I have not needed any specific clothing for this hobby. I would recommend just washing your hands up to your elbow before starting. In practice I didn't do this but had plenty of success after I got my SAB technique down.


Extras: Filter Print Post Top
OfflineSittin_On_A_Toilet
Stranger
Registered: 10/17/20
Posts: 16
Last seen: 3 years, 1 month
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041107 - 11/15/20 07:01 PM (3 years, 2 months ago)

I recently also went from B+ syringe to agar. Here are a couple posts I've made so far: https://www.shroomery.org/forums/showflat.php/Number/26989990
https://www.shroomery.org/forums/showflat.php/Number/27039803

Basically what you tried is the right way to do it, but if you don't have a good sterile technique down it will probably keep failing. Because you're in petri dishes you had to do a 'pour agar tek' right? Meaning you sterilize agar in a large bottle, then pour into petri dishes in a sterile environment. This step adds another chance of getting contam (although the contam on your plates looks like it came from your loop).

If you want to go as fool-proof as possible, try a No Pour tek using mason jars or a No Pour tek using little tupperware

Also keep in mind I've been at this for about a month total, so I am by no means an expert. But I had 9 out of 10 clean jars using the 'No Pour tek using mason jars' from a spore syringe, the 1 that contaminated I forgot the damn seal on the lid.... After those initial 9 clean jars, my spore syringe has sat in the fridge. If I want to start growth on another plate I just do agar to agar transfer.

Keep posting your results and lets follow each others first grows! Don't get discouraged, your first teks are supposed to fail. Learn from them, read more, and practice more!

Good luck!


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: CocaineBuffet]
    #27041119 - 11/15/20 07:09 PM (3 years, 2 months ago)

Quote:

CocaineBuffet said:
Personally I would go with the single drop and streak across the plate with the inoculation loop. Syringes just make the inoculation process convoluted. Most users here use prints/swabs.

Take things slow when working in the SAB. The more controlled and slow movements the less the air in the SAB gets stirred up.




if I got my hands on some sterile swabs the process would be:

1.flame syringe outside SAB, wipe needle and body of syringe with ISO once inside

2.Shoot syringe into sterile petri -cover it

3. open petri, swab, close it make an 'X' on clean agar plate

how many swipes can i get out of a swab? given my success rate how many plates would you make? 5-10?

I'm guessing I'll only need 1cc of spores then too. can I cap the needle and save it for my next attempt if this one fails? as long as I clean it up well with ISO?

Man, I thought I had a good bead on things, humbling.


Extras: Filter Print Post Top
OfflineCocaineBuffet
Stranger
 User Gallery


Registered: 08/29/19
Posts: 3,488
Last seen: 10 hours, 35 minutes
Trusted Cultivator
Re: My first grow, and I'm having some issues. [Re: Frozen Jedi]
    #27041125 - 11/15/20 07:14 PM (3 years, 2 months ago)

You are leaps and bounds ahead in knowledge for most green horn growers. The major hurdle is getting into agar which will become so simple and efficient you will beat yourself with a stick for not doing it sooner.

I would streak it the same way with that you have been doing if you are using a pre-sterilized swab such as this: https://www.amazon.com/Cotton-Tip-Applicator-Sterile-100/dp/B000BI3M60/ref=sr_1_2?dchild=1&keywords=sterilized+cotton+swab&qid=1605492805&sr=8-2

That is what I use for either applying prints to agar or taking swabs from fruits that don't traditionally drop spores.

2 to a pack so 1 plate for each. If you got your technique down you could do like 10+ with a single swab.


Extras: Filter Print Post Top
OfflineHikeadellic
Fungi Fan
Male


Registered: 08/31/20
Posts: 1,227
Loc: Appalachian Trail
Last seen: 12 days, 20 hours
Re: My first grow, and I'm having some issues. [Re: kanemush] * 2
    #27041127 - 11/15/20 07:18 PM (3 years, 2 months ago)

Maybe attempt grain to agar next time, clean up with a transfer before going back to grain


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: Sittin_On_A_Toilet]
    #27041143 - 11/15/20 07:36 PM (3 years, 2 months ago)

Quote:

Sittin_On_A_Toilet said:
I recently also went from B+ syringe to agar. Here are a couple posts I've made so far: https://www.shroomery.org/forums/showflat.php/Number/26989990
https://www.shroomery.org/forums/showflat.php/Number/27039803

Basically what you tried is the right way to do it, but if you don't have a good sterile technique down it will probably keep failing. Because you're in petri dishes you had to do a 'pour agar tek' right? Meaning you sterilize agar in a large bottle, then pour into petri dishes in a sterile environment. This step adds another chance of getting contam (although the contam on your plates looks like it came from your loop).

If you want to go as fool-proof as possible, try a No Pour tek using mason jars or a No Pour tek using little tupperware

Also keep in mind I've been at this for about a month total, so I am by no means an expert. But I had 9 out of 10 clean jars using the 'No Pour tek using mason jars' from a spore syringe, the 1 that contaminated I forgot the damn seal on the lid.... After those initial 9 clean jars, my spore syringe has sat in the fridge. If I want to start growth on another plate I just do agar to agar transfer.

Keep posting your results and lets follow each others first grows! Don't get discouraged, your first teks are supposed to fail. Learn from them, read more, and practice more!

Good luck!



Hey! looks like were on the same page, outstanding! stay in touch


Extras: Filter Print Post Top
OfflineFrozen Jedi
nooberist
Male

Registered: 10/17/20
Posts: 26
Last seen: 3 years, 9 days
Re: My first grow, and I'm having some issues. [Re: CocaineBuffet]
    #27041147 - 11/15/20 07:40 PM (3 years, 2 months ago)

Quote:

CocaineBuffet said:
You are leaps and bounds ahead in knowledge for most green horn growers. The major hurdle is getting into agar which will become so simple and efficient you will beat yourself with a stick for not doing it sooner.

I would streak it the same way with that you have been doing if you are using a pre-sterilized swab such as this: https://www.amazon.com/Cotton-Tip-Applicator-Sterile-100/dp/B000BI3M60/ref=sr_1_2?dchild=1&keywords=sterilized+cotton+swab&qid=1605492805&sr=8-2

That is what I use for either applying prints to agar or taking swabs from fruits that don't traditionally drop spores.

2 to a pack so 1 plate for each. If you got your technique down you could do like 10+ with a single swab.



awesome info! thanks for the boost man, I haven't got my ass handed to me like this in a long time. It's fuckin fun though!


Extras: Filter Print Post Top
Jump to top Pages: 1 | 2 | Next >  [ show all ]

Shop: Mushroom-Hut Liquid Cultures   PhytoExtractum Buy Bali Kratom Powder   North Spore Injection Grain Bag   Left Coast Kratom Buy Kratom Extract   Original Sensible Seeds Bulk Cannabis Seeds


Similar ThreadsPosterViewsRepliesLast post
* Grow bags slow/stalling braxton08 103 3 11/21/20 01:02 PM
by Hikeadellic
* transferring mycelium wikedanjel 4,117 13 01/10/03 03:54 AM
by debianlinux
* Re: DB Issue (Pic) Holydiver 1,510 15 04/03/01 01:03 PM
by Ryche Hawk
* Bulk Growing AlphaWave 4,734 3 10/22/01 05:02 PM
by phrozendata
* 1st time grow 3ozs dry onlygrowgiants 2,618 7 08/21/02 10:58 AM
by _VisioN_
* Mex A Grain-to-Grain Transfers babyshroom 1,956 11 08/29/02 08:05 AM
by babyshroom
* Questions on growing P. Cyanescens... Wysefool 2,702 5 01/06/03 01:17 PM
by hermes3
* Re: Why do I want to grow mycelium on Agar?? Anonymous 2,476 3 02/13/00 04:07 PM
by AnubisRonin

Extra information
You cannot start new topics / You cannot reply to topics
HTML is disabled / BBCode is enabled
Moderator: Shroomism, george castanza, RogerRabbit, veggie, mushboy, fahtster, LogicaL Chaos, 13shrooms, Stipe-n Cap, Pastywhyte, bodhisatta, Tormato, Land Trout, A.k.a
349 topic views. 28 members, 151 guests and 28 web crawlers are browsing this forum.
[ Show Images Only | Sort by Score | Print Topic ]
Search this thread:

Copyright 1997-2024 Mind Media. Some rights reserved.

Generated in 0.022 seconds spending 0.007 seconds on 14 queries.