Liquid Culture Fresh Air Exchange

p2k1 · 04-05-09, 23:39

Hey everyone a FOAF recently prepared 8 liquid cultures of 250ml water to a tea spoon of natural honey, 7 were microwaved on high for 5 minutes, 1 was pressure cooked for about 20 minutes at 15 psi. The containers used were not jars but Ziploc Twist n Loc [image]http://3.bp.blogspot.com/_gTvuTXdszC4/SEXu6lBZC7I/AAAAAAAABsM/KaKoH7EgvBo/s320/ziploc+twist.jpg[/image] with a layer of clingwrap seperating the container from the lid for the purposes of keeping contaminants out of the LC when the needles going in or out. Of the 8 only one shows significant mycellium growth (not the PCed one). After injecting the LCs and two additional jars there was a fraction of a CC remaining in the syringe so it was filled with lc from one of the sterilized containers and shows growth of myc in the syringe, a great tip picked up from someone here. Bear in mind these were made roughly a week ago, perhaps a bit less. Two test jars of rye were inoculated with 1cc each of the remaining multispore and only one has shown minor growth so far.
Anyways these twist n loc containers don't have holes in the lids. They have been untightened slightly so that they are not sealed. Should a hole be made in these lids and be covered with micropore tape?
Any advice is greatly appreciated.
Stay free

golly · 04-06-09, 00:20

You don't want to loosen any lid on a LC after they have cooled..If there is at least a 1/4pint of air trapped, then that should be plenty to grow some Myc..
You may have to figure a way to relieve the vacuum when you go to suck the goods out
though..

Mushrooms201 · 04-06-09, 01:46

I have temporarily given up on LCs for the moment after fighting with that question. I seemed to get stronger growth with an air exchange port when i was doing it or I also would sterilize an empty jar to darw from or inject clean air into the culture not sure if it ever did any good though. For now I will stick to agar and grain to grain.

GreenBastard · 04-06-09, 22:57

Quote:

Originally Posted by golly

You don't want to loosen any lid on a LC after they have cooled..If there is at least a 1/4pint of air trapped, then that should be plenty to grow some Myc..
You may have to figure a way to relieve the vacuum when you go to suck the goods out
though..

Would an airport inserted into the injection be enough to let in some fresh air and alleviate any sort of vacuum?

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04-05-09, 23:39	#1 (permalink)
p2k1 Mycotopiate Join Date: Jul 1972 Posts: 326	Liquid Culture Fresh Air Exchange Hey everyone a FOAF recently prepared 8 liquid cultures of 250ml water to a tea spoon of natural honey, 7 were microwaved on high for 5 minutes, 1 was pressure cooked for about 20 minutes at 15 psi. The containers used were not jars but Ziploc Twist n Loc [image]http://3.bp.blogspot.com/_gTvuTXdszC4/SEXu6lBZC7I/AAAAAAAABsM/KaKoH7EgvBo/s320/ziploc+twist.jpg[/image] with a layer of clingwrap seperating the container from the lid for the purposes of keeping contaminants out of the LC when the needles going in or out. Of the 8 only one shows significant mycellium growth (not the PCed one). After injecting the LCs and two additional jars there was a fraction of a CC remaining in the syringe so it was filled with lc from one of the sterilized containers and shows growth of myc in the syringe, a great tip picked up from someone here. Bear in mind these were made roughly a week ago, perhaps a bit less. Two test jars of rye were inoculated with 1cc each of the remaining multispore and only one has shown minor growth so far. Anyways these twist n loc containers don't have holes in the lids. They have been untightened slightly so that they are not sealed. Should a hole be made in these lids and be covered with micropore tape? Any advice is greatly appreciated. Stay free

04-06-09, 01:46	#3 (permalink)
Mushrooms201 Mycotopiate Join Date: Mar 2009 Posts: 541	Same question I have temporarily given up on LCs for the moment after fighting with that question. I seemed to get stronger growth with an air exchange port when i was doing it or I also would sterilize an empty jar to darw from or inject clean air into the culture not sure if it ever did any good though. For now I will stick to agar and grain to grain.

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04-06-09, 00:20	#2 (permalink)
golly modapotato Join Date: Oct 2005 Posts: 6,198	You don't want to loosen any lid on a LC after they have cooled..If there is at least a 1/4pint of air trapped, then that should be plenty to grow some Myc.. You may have to figure a way to relieve the vacuum when you go to suck the goods out though..