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OfflineSma11mau5
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Registered: 08/15/12
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2 batches from same nocc...only one is growning help/advice needed
    #16830385 - 09/11/12 05:38 PM (12 years, 4 months ago)

Hoping to get some questions answered regarding a situation in which 8 jars were nocc'd in the same day from the same pc batch with the same style lids the only diff is 4 were galindoi and 4 were tampanesis, each jar got approx 2.5 ml of MS from spore works :syringe:



Now.. The galindoi are GREAT, but the tampanesis never did anything ... at all ... no contam ... no mycelium either ... So what is the best course of action??

-turf the tampanesis jars and redo them with new fresh grain and a fresh syringe of either Mexican a or jalisco(RR grain tek with some of stonesun's suggestions)

or

- treat them like a purchaced sterile grain jar and simply reinoculate with either g2g from the nice galindoi's or a new MS syringe

or

- wait longer...keep in mind since those pics were taken the galindoi has almost completely completed visible colonization and the tamps show NOTHING happening at all, not even the slight condensation seen on jars that have any activity






Why do you think this might have happened?? My reasoning is that the only thing that could have caused it other than simply a dead syringe ...mayyyybe..is that viable spores got killed during flame sterilizing, but it seems unlikely considering I changed no method for the galindoi and they turned out great...has anyone had his happen before, had anyone gotten refunds from the works for a dead MS?

Any help would be appreciated :thumbup:

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OfflineTheDeathryder
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16830398 - 09/11/12 05:41 PM (12 years, 4 months ago)

How long ago did you inoculate?


Edit:  Nevermind, I see the date on the label now.  I would contact the vendor and tell them that the spores were non viable for microscopy purposes, and request another syringe.

Edited by TheDeathryder (09/11/12 05:43 PM)

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OfflineCosmic Fool
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16830421 - 09/11/12 05:45 PM (12 years, 4 months ago)

Blank shots are a fairly common occurrance with ms syringes.  Aside from getting a replacement, I would just try to reinnoculate the jars, since it sounds like there might not be a massive sporeload in that syringe and I wouldn't consider it worth putting more materials on the line for a syringe that might not even work.  I would attempt to get an lc growing from that syringe and use that in the future for jars.

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OfflineSma11mau5
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Cosmic Fool]
    #16830501 - 09/11/12 06:02 PM (12 years, 4 months ago)

Quote:

TheDeathryder said:
How long ago did you inoculate?


Edit:  Nevermind, I see the date on the label now.  I would contact the vendor and tell them that the spores were non viable for microscopy purposes, and request another syringe.




Will do! Thanks for the always quick response man!

Quote:

Cosmic Fool said:
Blank shots are a fairly common occurrance with ms syringes.  Aside from getting a replacement, I would just try to reinnoculate the jars, since it sounds like there might not be a massive sporeload in that syringe and I wouldn't consider it worth putting more materials on the line for a syringe that might not even work.  I would attempt to get an lc growing from that syringe and use that in the future for jars.




Unfortunatly there is no more tampanesis MS left so reinoculateing would have to be with either the mex a or Jalisco ...if it was between doing one of those varietys or sacrificing a galindoi jar to a G2G transfer what would you reccomend?

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OfflineTheDeathryder
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16830528 - 09/11/12 06:09 PM (12 years, 4 months ago)

G2G would colonize faster, so that would be my pick.  Of course I'm also a fan of starting spores on agar, so I'm biased on that.

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OfflineSma11mau5
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: TheDeathryder]
    #16830555 - 09/11/12 06:13 PM (12 years, 4 months ago)

agar and a stack was added to the supplies yesterday, it's currently cooling right now...would you say there are any worries about PCing the dishes? If they are in the sleeve and I don't open till its in the glove box should it be ok? And then what about storage and possible reusing if the dishes,...sorry for all the questions, just stoked to start studying agar :grin:

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OfflineTheDeathryder
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16830580 - 09/11/12 06:17 PM (12 years, 4 months ago)

The only dishes I've worked with were prepoured.  I don't have access to a flowhood, so I'm a fan of no pour agar jars on that front.  I would store agar dishes in the fridge until your ready to use them, if they're glass, they should be reusable, I have read about plastic dishes melting, so I would be wary on that front.

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OfflineCosmic Fool
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: TheDeathryder]
    #16830899 - 09/11/12 07:14 PM (12 years, 4 months ago)

Are you talking about a sleeve of plastic petri dishes, sealed? If so, you're not supposed to be PC those, and they're sterile to begin with anyway.  You can pour the dishes in a glovebox no problem.  Just PC the agar in a whiskey bottle or something similar and pour the dishes before the solution cools and solidifies.

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Invisibletahoe
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Cosmic Fool]
    #16830927 - 09/11/12 07:18 PM (12 years, 4 months ago)

Looks like a bad syringe. It does happen to the best of them. Contact sw, get a new one.
DO NOT REUSE JARS!! You can pc them again as is and use them but do not just add more spores. You are not 100% sure of what is going on inside the jar.


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


My Legacy
https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987

Teh=The
I need to proofread

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Offlinearkady2300
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: tahoe]
    #16831429 - 09/11/12 08:26 PM (12 years, 4 months ago)

Great thread about saving a otherwise awful situation.

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OfflineSma11mau5
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: tahoe]
    #16832202 - 09/11/12 10:30 PM (12 years, 4 months ago)

Quote:

tahoe said:
Looks like a bad syringe. It does happen to the best of them. Contact sw, get a new one.
DO NOT REUSE JARS!! You can pc them again as is and use them but do not just add more spores. You are not 100% sure of what is going on inside the jar.




So to understand you-before they are used again, a quick run though the PC and then inoculate as usual... Easy enough :thumbup: at least it saves needing to soak/prep more grain :rockon:

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OfflineSma11mau5
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16832229 - 09/11/12 10:34 PM (12 years, 4 months ago)

Another question came to mind...could you wait until the sclerotia harvest, and then instead of casing and fruiting the remaining picked-over-spawn just using it as a grain master to fire up your next batch of jars... Might have the danger to be contam city but if kept clean it could be sweet for sclerotia fans as an easy easy way to always start their next batch, thoughts??

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OfflineCosmic Fool
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Registered: 09/07/12
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16832243 - 09/11/12 10:36 PM (12 years, 4 months ago)

Don't harvest the jars and *then* use as grain master.  That's just a recipe for contams.  If you want to do grain transfers, do that and then harvest the resulting sclerotia from those subsequent jars when they are finished.

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Offlinezepski
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16832251 - 09/11/12 10:37 PM (12 years, 4 months ago)

Quote:

Sma11mau5 said:
Another question came to mind...could you wait until the sclerotia harvest, and then instead of casing and fruiting the remaining picked-over-spawn just using it as a grain master to fire up your next batch of jars... Might have the danger to be contam city but if kept clean it could be sweet for sclerotia fans as an easy easy way to always start their next batch, thoughts??




I dont know the answer here i've not much experience with sclerotia.  I've got my 1st set of jars going now for a few months. 

But to make an adjustment on the idea of g2g.  Could you make a GLC from the jar and proceed from there?

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OfflineCosmic Fool
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: zepski]
    #16832260 - 09/11/12 10:40 PM (12 years, 4 months ago)

Yes indeed op, you could use a syringe to squirt some sterile water in to one of the fully colonized jars (be sure it is free of any contams!), then suck it back up and use that as inoculant to produce more lc, or more jars, whatever.  Then you can have it both ways by harvesting your sclerotia as well as propagating more jars.

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Invisibletahoe
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16833011 - 09/12/12 01:47 AM (12 years, 4 months ago)

Quote:

Sma11mau5 said:
Another question came to mind...could you wait until the sclerotia harvest, and then instead of casing and fruiting the remaining picked-over-spawn just using it as a grain master to fire up your next batch of jars... Might have the danger to be contam city but if kept clean it could be sweet for sclerotia fans as an easy easy way to always start their next batch, thoughts??



You know in theory you could. If said jar stayed sterile as fuck, if you were sterile when you removed the sclerotia. Main concern would be metabolites. One usually gets some built up while waiting for sclerotia. best bet is to do your grain masters as soon as they are fully colonized. Do not try this unless you are experienced. Its much different that squirting some spores in a jar or dropping an agar chunk in there.

Yes to reusing the jars that show no sign of life. As long as they look clean as can be. Just pc them again, cool them and go for it. Moisture might be a little whacked but no where near enough to cause a major problem.


--------------------
Stop experimenting half way through your first grow. Grow it to maturity, watch it, learn from it. Do this a few times then experiment with different ideas and figure out what works best for you.


My Legacy
https://www.shroomery.org/forums/showflat.php/Number/22140987#22140987

Teh=The
I need to proofread

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OfflineSma11mau5
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Registered: 08/15/12
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: tahoe]
    #16833860 - 09/12/12 09:44 AM (12 years, 4 months ago)

It's already referred to in another thread but do you think it would be safe to redo the lids with the new ez felt filter discs that are being made or just not even open them and simply re-PC without screwing around, the only thing I'm most worried about is messing up moisture levels by having the lids off for the time it takes for the silicone to dry...or could just throw unaltered plastic lids on for the wait period?

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OfflineSma11mau5
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Cosmic Fool]
    #16833942 - 09/12/12 10:10 AM (12 years, 4 months ago)

Quote:

Cosmic Fool said:
Don't harvest the jars and *then* use as grain master.  That's just a recipe for contams.  If you want to do grain transfers, do that and then harvest the resulting sclerotia from those subsequent jars when they are finished.




Got it :smile:


Quote:

zepski said:
Quote:

Sma11mau5 said:
Another question came to mind...could you wait until the sclerotia harvest, and then instead of casing and fruiting the remaining picked-over-spawn just using it as a grain master to fire up your next batch of jars... Might have the danger to be contam city but if kept clean it could be sweet for sclerotia fans as an easy easy way to always start their next batch, thoughts??




I dont know the answer here i've not much experience with sclerotia.  I've got my 1st set of jars going now for a few months. 

But to make an adjustment on the idea of g2g.  Could you make a GLC from the jar and proceed from there?



Quote:

Cosmic Fool said:
Yes indeed op, you could use a syringe to squirt some sterile water in to one of the fully colonized jars (be sure it is free of any contams!), then suck it back up and use that as inoculant to produce more lc, or more jars, whatever.  Then you can have it both ways by harvesting your sclerotia as well as propagating more jars.




This seems like the absolute best plan!! What are the dangers though, could the tiny amount of excess sterile water cause a problem with the moisture content of an otherwise healthy jar or is the worry small enough to just go ahead and risk it?

If this were done, the method would be to fill a syringe with Reverse osmosis water(no access to distilled, so we get culligan drinking water in the 10 gallon bottles) , wrap in tinfoil and PC ( but for how long?) and then transfer the syringe and the grain jar to be used into the glovebox...then proceed as above? Thoughts?

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OfflineCosmic Fool
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Registered: 09/07/12
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Re: 2 batches from same nocc...only one is growning help/advice needed [Re: Sma11mau5]
    #16834474 - 09/12/12 12:12 PM (12 years, 4 months ago)

Quote:

could the tiny amount of excess sterile water cause a problem with the moisture content of an otherwise healthy jar or is the worry small enough to just go ahead and risk it?




Try to get as much water out of the jar as you can, to avoid leaving excess.  After that, you the jar can be spawned to bulk if that's what you want to do.  A few drops of water isn't going to mess anything up.

Quote:

If this were done, the method would be to fill a syringe with Reverse osmosis water(no access to distilled, so we get culligan drinking water in the 10 gallon bottles)




Sounds fine.  You don't have to use reverse osmosis though, tap will work fine.

Quote:

wrap in tinfoil and PC ( but for how long?) and then transfer the syringe and the grain jar to be used into the glovebox...then proceed as above? Thoughts?




Yes, make sure the syringe is upside down (needle pointing up) while pcing, or else the pressure will force much of the water out.  It doesn't have to be totally vertical, just get it as up as you can.  Then proceed as above, yes.

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