|
Some of these posts are very old and might contain outdated information. You may wish to search for newer posts instead.
|
mikesethnobotany
Man
Registered: 07/29/11
Posts: 1,267
Loc: Missouri, USA
Last seen: 2 years, 8 months
|
Re: Shiitake Mycelium [Re: NSF]
#15234294 - 10/16/11 03:38 PM (12 years, 5 months ago) |
|
|
Heres another update with my shiitake on grain. The proof is in the pudding. The rye grain is now colonizing well after being exposed to natural light for just a few days. Growth is much thicker and appears to be spreading much faster than before when I had the jars in the dark.
To those of you (EM) who have been worried that moisture of the rye grain could have been the culprit.... you're truthfully 50% right. After some time of thinking about the issue I realized something....
About a week before inoculating these jars with agar, I used my spore syringe to try and do L2G. This grew very slowly in my dark cupboard (light is the other 50% of the porblem, well maybe slightly more like 65%) so I just gave up and said "Im gunna put agar in there."
So long story short, the jars had an extra week on them that they had been exchanging gas -- and therefor moisture -- which has probably been sufficient enough to dry the grain out, like some of you speculated. I am astonished of this discovery and how on the spot you guys have been about it the entire time.... Thanks so much for your help guys! I will be less reluctant to take your advice immediately in the future... sometimes I amaze myself!
Good news however is that these two jars are growing quickly..
Now, to figure out my slant predicament
|
EvilMushroom666
Heretic
Registered: 11/18/09
Posts: 10,289
Loc: Canada
|
|
Glad to see things are moving along....
I saw your slant post...really odd that it has taken that long to grow out, have you capped your slants tight? You want some gas exchange to take place, perhaps that is the issue?
When using cultubes by Simport I leave the cap in the open position and wrap with parafilm before letting it grow out.
With glass slants and screw tops I screw the caps down tight and then back it off around half a turn and wrap the gap between the cap and slants with parafilm.
|
mikesethnobotany
Man
Registered: 07/29/11
Posts: 1,267
Loc: Missouri, USA
Last seen: 2 years, 8 months
|
|
Hmmm now that you mention it there is absolutely no gas exchange with these slants because I am using the glass serum vacutainers. Alrighty, this makes sense - I will be pulling the caps up so that the miniature gas exchange crevice in the vacutainer lid can transport gas... although this still will not be much gas exchange with the parafilm covering it.... maybe it would be more beneficial if I just took the tops of and just capped the vacutainers solely with parafilm?
|
EvilMushroom666
Heretic
Registered: 11/18/09
Posts: 10,289
Loc: Canada
|
|
Simport cultubes have two positions you can set the lid, anaerobic and aerobic. After I am done slanting a culture I place the lid in its aerobic position and wrap the tubes as shown below:
I would first try to lift your lid off a bit and allow the slots in the tubes(I assume you are using vacutainers) to be exposed and wrap with parafilm. Obviously do this under aseptic conditions.
In the past I have used parafilm to wrap the entire end of a culture tube and it worked fine, but this is obviously not the optimal way to go about things for long term storage.
|
mikesethnobotany
Man
Registered: 07/29/11
Posts: 1,267
Loc: Missouri, USA
Last seen: 2 years, 8 months
|
|
Quote:
In the past I have used parafilm to wrap the entire end of a culture tube and it worked fine, but this is obviously not the optimal way to go about things for long term storage.
Right. I was thinking I could do this, then once the culture reaches 70-80% of the slant then recap for storage in the fridge. But still, that doesn't leave out the possibility that leaving the culture to having no gas exchange in the fridge would result in a bad culture over time... still not sure how I feel about this. I think that in the future I will be purchasing these cultubes you are using... seems like all the pros are using them these days
|
EvilMushroom666
Heretic
Registered: 11/18/09
Posts: 10,289
Loc: Canada
|
|
My understanding is starving mycelium of oxygen will lead to eventual death of the culture.
I recommend Simports 10cc culture tubes, or any glass screw top vials with a large enough opening (around 16mm).
|
DRCola
Registered: 05/04/11
Posts: 134
Loc: USA
|
|
Hope this post isn't too old to "bump".
But I'm so glad I found and read this post. My Shiitake grain jars are basking in the indirect sunlight as a result. I inoculated 3 days ago and don't yet see anything, but they just went into the light.
It's my first Shiitake to grain. It's from a vendor sponsor's syringe. I was thinking of contacting them until I read this post.
It leads me to another question. What could one do about grain that may be too dry? I prepared the rye grain jars a week before inoculation.
Thanks all.
|
|