|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
Wolfgang187
Stranger



Registered: 12/15/13
Posts: 33
Last seen: 9 years, 8 months
|
Beginner cant get jars to colonize
#19278719 - 12/15/13 10:49 AM (10 years, 2 months ago) |
|
|
So, I'm, a complete newb at this, but I followed the instructions of this wbs tek which has been suggested to be foolproof: http://www.shroomery.org/9030/Doc34-s-Wild-Bird-Seed-Tek
I followed the tek 100% to the letter.
1st attempt I incubated after inoculation @ 80 degrees in a TIT. Not one jar colonized after waiting 2 months.
2nd attempt I only incubated 1/2 the jars and left the other half out at room temperature. Same result after 2 months.
24 jars, 24 complete failures without a single jar colonizing even a little tiny bit. At this point I'd be happy with colonized jars full of contaminants. At least there'd be life in them.
Is there a tek even more foolproof than the one I listed? At this point I'd happily accept a tek that usually results in contamination 2/3 of the time. It would be better than where I am now.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
|
Re: Beginner cant get jars to colonize [Re: Wolfgang187]
#19278722 - 12/15/13 10:50 AM (10 years, 2 months ago) |
|
|
agar teks are more fool proof
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
|
Re: Beginner cant get jars to colonize [Re: cronicr]
#19278772 - 12/15/13 11:03 AM (10 years, 2 months ago) |
|
|
That's rough man. All I can think is either the spors you are using arnt viable or you are trying to inoculate while your jars are still to hot from PCing
|
Wolfgang187
Stranger



Registered: 12/15/13
Posts: 33
Last seen: 9 years, 8 months
|
Re: Beginner cant get jars to colonize [Re: MudaFuka]
#19278775 - 12/15/13 11:04 AM (10 years, 2 months ago) |
|
|
Quote:
MudaFuka said: That's rough man. All I can think is either the spors you are using arnt viable or you are trying to inoculate while your jars are still to hot from PCing
I'm letting the jars cool overnight
|
Josh.0
ConnoissurOfSorts


Registered: 11/25/13
Posts: 553
|
Re: Beginner cant get jars to colonize [Re: Wolfgang187]
#19278786 - 12/15/13 11:07 AM (10 years, 2 months ago) |
|
|
are you letting your needle cool after flaming it,before you inoculate?
|
monoculture
Mycaddict



Registered: 10/16/13
Posts: 246
Last seen: 9 years, 6 months
|
Re: Beginner cant get jars to colonize [Re: Wolfgang187]
#19278802 - 12/15/13 11:11 AM (10 years, 2 months ago) |
|
|
Post pictures of the process, let us know what you inoculate them with. With more info we might be able to help you find the cause of your failure. Now it's just a wild guess.
If he wouldn't let the needle cool down before inoculation, only the first part of the solution would be sterile, there should be viable spores when injecting 1cc even when immediately hitting when needle is still red.
I have the idea your spore solution is no good.
Edited by monoculture (12/15/13 11:14 AM)
|
Wolfgang187
Stranger



Registered: 12/15/13
Posts: 33
Last seen: 9 years, 8 months
|
Re: Beginner cant get jars to colonize [Re: Josh.0]
#19278814 - 12/15/13 11:13 AM (10 years, 2 months ago) |
|
|
Quote:
Josh.0 said:
are you letting your needle cool after flaming it,before you inoculate?
Yes. 1/2 the jars I didn't even flame sterilize to see if that was my problem. No matter how I sterilize, no life whatsoever.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
|
Re: Beginner cant get jars to colonize [Re: Wolfgang187]
#19278821 - 12/15/13 11:13 AM (10 years, 2 months ago) |
|
|
sounds like you need new spores
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
|
Re: Beginner cant get jars to colonize [Re: MudaFuka]
#19278829 - 12/15/13 11:15 AM (10 years, 2 months ago) |
|
|
One more thing I just looked at the tek you used and it should work. I would just change a few minor details. 1.Soak for 24 hours not 14 2.PC for 90 minutes not 60 3. Don't use poste ofice tyvek. Use tyvek from disposable painter's coveralls.
Other than that follow the tek and you should be fine.
|
Wolfgang187
Stranger



Registered: 12/15/13
Posts: 33
Last seen: 9 years, 8 months
|
Re: Beginner cant get jars to colonize [Re: MudaFuka]
#19278882 - 12/15/13 11:24 AM (10 years, 2 months ago) |
|
|
Quote:
MudaFuka said: One more thing I just looked at the tek you used and it should work. I would just change a few minor details. 1.Soak for 24 hours not 14 2.PC for 90 minutes not 60 3. Don't use poste ofice tyvek. Use tyvek from disposable painter's coveralls.
Other than that follow the tek and you should be fine.
Youre telling me that those things could cause complete failure to colonize... 24 times?
|
SuperSillyUs
Proud 'Shitbird'. ®

Registered: 12/14/13
Posts: 326
Last seen: 10 years, 2 months
|
Re: Beginner cant get jars to colonize [Re: MudaFuka]
#19278909 - 12/15/13 11:30 AM (10 years, 2 months ago) |
|
|
Sounds like you'll 'need new spores'...one way or the other. If you're going to try again.* Take heart...Some people try (and fail) half a dozen times until some random seeming variable is changed (no/new carpet in next apartment).
Contamination would cause all your jars to stall. Or in this case not do anything. Too dry sub, too wet. Following a tek to the tee, still leaves that elusive 'grower's eye' ...or 'grower's feel' that has to be developed over years. Maddening, isn't it?
Keep trying. I think usually when there is zero growth, that's a sign of contamination? Or something. What sterile technique did you use?
The tek you linked to doesn't mention a still air box or flow hood at all. Does it? I just glanced over it quickly...
Same vendor? Same sterile technique? How far apart in time were the attempts?
Also...what is a TIT? Why is that advantageous? Oh ...I see...really? People still use that?
And, I'm sure the group will correct me if I';m wrong but, 80 degrees seems a little bit warm, right? No?
...not sure what the consensus is on that last one.
* I wouldn't be quick to blame the vendor. Especially if this was your first (and second) attempt.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
|
Re: Beginner cant get jars to colonize [Re: cronicr]
#19278920 - 12/15/13 11:31 AM (10 years, 2 months ago) |
|
|
Quote:
cronicr said: agar teks are more fool proof
it's the best way to germinate spores and guarentee these problems can be avoided
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
invitro


Registered: 05/03/13
Posts: 2,529
Last seen: 2 months, 14 days
|
Re: Beginner cant get jars to colonize [Re: cronicr]
#19278951 - 12/15/13 11:40 AM (10 years, 2 months ago) |
|
|
How about some pics of the jars? It could be the syringe was shipped to you during a time when temps were really high and killed the spores. Imagine your package in the cargo compartment of a plane sitting on a tarmac for 24 hours on a hot sunny day because the engine suddenly needed repair and the shipment was delayed. High temperatures might have cooked your syringe. Try again. I'd ditch the tit incubator unless you have strict control of temp fluctuations in your house. Sudden and unnoticed temp spikes encourage contams to grow and hurt the myc. I wouldn't soak for 24 hours either.
|
Stargaze
Stranger



Registered: 11/11/13
Posts: 120
Loc: Sporeville
Last seen: 9 years, 9 months
|
Re: Beginner cant get jars to colonize [Re: invitro]
#19279061 - 12/15/13 12:06 PM (10 years, 2 months ago) |
|
|
Where did you get your spores from (out of curiosity)
--------------------
|
MudaFuka
Poppin bottles



Registered: 12/14/13
Posts: 18,648
Loc: Canada
|
Re: Beginner cant get jars to colonize [Re: Wolfgang187]
#19279214 - 12/15/13 12:40 PM (10 years, 2 months ago) |
|
|
Quote:
Wolfgang187 said:
Quote:
MudaFuka said: One more thing I just looked at the tek you used and it should work. I would just change a few minor details. 1.Soak for 24 hours not 14 2.PC for 90 minutes not 60 3. Don't use poste ofice tyvek. Use tyvek from disposable painter's coveralls.
Other than that follow the tek and you should be fine.
Youre telling me that those things could cause complete failure to colonize... 24 times?
No I would say spores are the problemb I was simply pointing out things I would do difrently next time
|
SuperSillyUs
Proud 'Shitbird'. ®

Registered: 12/14/13
Posts: 326
Last seen: 10 years, 2 months
|
Re: Beginner cant get jars to colonize [Re: MudaFuka]
#19279245 - 12/15/13 12:46 PM (10 years, 2 months ago) |
|
|
Wouldn't having tubs of water sitting around for weeks be a breeding ground for bacteria and contamination?
|
Wolfgang187
Stranger



Registered: 12/15/13
Posts: 33
Last seen: 9 years, 8 months
|
Re: Beginner cant get jars to colonize [Re: SuperSillyUs]
#19279526 - 12/15/13 01:56 PM (10 years, 2 months ago) |
|
|
Quote:
SuperSillyUs said: Wouldn't having tubs of water sitting around for weeks be a breeding ground for bacteria and contamination?
I added a bit of bleach to the water. Even still, I left half of the jars outside of the incubator with the same result.
|
Juicin
Stranger

Registered: 10/27/13
Posts: 897
|
Re: Beginner cant get jars to colonize [Re: Wolfgang187]
#19279559 - 12/15/13 02:04 PM (10 years, 2 months ago) |
|
|
I'd get new genetics OP. As others have said agar is the best way to tell but it's so cheap you might as well order now and try your old shit on something you can see what's going on.
|
Wolfgang187
Stranger



Registered: 12/15/13
Posts: 33
Last seen: 9 years, 8 months
|
Re: Beginner cant get jars to colonize [Re: Stargaze]
#19280138 - 12/15/13 04:39 PM (10 years, 2 months ago) |
|
|
Quote:
Stargaze said: Where did you get your spores from (out of curiosity)
dunno if I'm allowed to post where I got my spores from, but they weren't from a sponsor of this site. I think I recall someone saying that you should be able to see the spores floating around in a syringe, but from what I remember my syringes were pretty clear.
|
cronicr



Registered: 08/07/11
Posts: 61,436
Loc: Van Isle
Last seen: 2 years, 1 month
|
Re: Beginner cant get jars to colonize [Re: Wolfgang187]
#19280157 - 12/15/13 04:42 PM (10 years, 2 months ago) |
|
|
spore are micro scopic so you don't "need" to see them in order for them to be there, i prefer mine to be on the clear side
--------------------
  It doesn't matter what i think of you...all that matters is clean spawn I'm tired do me a favor
|
|