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blthirteenre



Registered: 06/10/12
Posts: 430
Loc: New York
Last seen: 9 months, 17 days
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QUESTION ABOUT INOCULATING W/ AGAR (PICS)
#18736918 - 08/21/13 11:01 AM (10 years, 5 months ago) |
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i have been using liquid culture for the inoculation of grain and pf cakes for the past couple of years, and after a contamination catastrophe with my liquid culture, contaminating about 100 pf cakes, i decided to figure out agar. i also believe i quit pf cakes.
anyway, i first tried doing tissue cloning. i failed 100%. i may have not been making the dishes correctly, but i decided to put a few drops of a vendor's syringe (and syringes i made with spore prints) onto the agar dish which turned out to be much better.
my question is: can i inoculate a rye grain jar from a petri dish that has some sort of contamination on it, or is it too risky? i used these dishes:

and the jars turned out contaminated.
i have this one dish that turned out VERY beautiful:

but i just noticed there is some contamination on the lid [see: 4 o'clock]! these dishes came in not mint condition, i got 60 for $15 free s/h and the bags were not in good condition (the guy sent me a free sleeve). they are also much smaller than i expected. i think buying dishes on ebay is very risky; i am pretty alot of the contamintion has to do with the dishes not being perfectly sterile.
anyway, i would love to use this agar but is it inevitable that i will contaminate my jars?
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Trippy_Penguin



Registered: 03/18/12
Posts: 624
Last seen: 3 years, 5 months
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: blthirteenre]
#18736942 - 08/21/13 11:05 AM (10 years, 5 months ago) |
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I'd transfer away from the contaminants before I attempted a transfer to grain.
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SpitballJedi
Ancient Astronaut


Registered: 10/13/12
Posts: 8,598
Loc: Nibiru
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: Trippy_Penguin]
#18736959 - 08/21/13 11:10 AM (10 years, 5 months ago) |
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Part of the reason for using agar is so you can get a nice clean culture. Don't waste your time and grains with a known contaminated culture.
Cut a piece of the clean myc out and transfer it to a new agar dish. Do this until you at least have no contams.
You can also transfer till you have an isolate.
-------------------- The Basics A little civility goes a long way The Noob Forum The Hammock Hangers' Forum
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FrankHorrigan
The Inquisition



Registered: 01/04/11
Posts: 10,573
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: Trippy_Penguin]
#18736978 - 08/21/13 11:15 AM (10 years, 5 months ago) |
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Quote:
petersodm said: I'd transfer away from the contaminants before I attempted a transfer to grain.

In fact, I sometimes take it a step further and take one more transfer from the healthy plate as well.
My reasoning is I want no contaminants on the plate, nor any mycelium that originated from that contaminated dish.
Once you see green in the petri, you can assume it is literally vomiting spores all over the mycelium. They are just waiting for a nice home to germinate in, such as the grains.
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blthirteenre



Registered: 06/10/12
Posts: 430
Loc: New York
Last seen: 9 months, 17 days
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: SpitballJedi]
#18736986 - 08/21/13 11:17 AM (10 years, 5 months ago) |
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altho there is a contaminate ,is that last dish an isolate? is there anymore sectoring to do?
yeah, you guys are right i suppose, it is just that i am pretty confident that i have been buying non sterile dishes and clean agar dishes aren't something i have this moment. a lack of patience will certainly be the death of me. better safe than sorry, i suppose.
thanks!
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FrankHorrigan
The Inquisition



Registered: 01/04/11
Posts: 10,573
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: blthirteenre]
#18736990 - 08/21/13 11:19 AM (10 years, 5 months ago) |
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Quote:
blthirteenre said: altho there is a contaminate ,is that last dish an isolate?
I see at least 4 sectors on that just looking at it from the top.
How many transfers from MS is that dish?
You will not see good sectoring with cubes until usually the 4th, 5th transfers.
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blthirteenre



Registered: 06/10/12
Posts: 430
Loc: New York
Last seen: 9 months, 17 days
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: FrankHorrigan]
#18737015 - 08/21/13 11:28 AM (10 years, 5 months ago) |
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Quote:
FrankHorrigan said:
Quote:
blthirteenre said: altho there is a contaminate ,is that last dish an isolate?
I see at least 4 sectors on that just looking at it from the top.
How many transfers from MS is that dish?
You will not see good sectoring with cubes until usually the 4th, 5th transfers.
that is the first transfer. i did a few drops of the syringe onto dish #1, then transfered it THAT dish: dish #2.
damn, atleast 4 sectors? that is crazy. i was reading someone's guide to doing agar ,i forget his name but it is his "thing", and my eye is still imperfect when it comes to sectoring. i guess i will aim for that think portion of myc between 3-6 o'clock.
frankhorrigan: what size dishes do you prefer? these are 50 or 60mmx15mm
can someone provide me with a reliable petri dish source? 100% sterile?
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FrankHorrigan
The Inquisition



Registered: 01/04/11
Posts: 10,573
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: blthirteenre]
#18737030 - 08/21/13 11:33 AM (10 years, 5 months ago) |
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Quote:
blthirteenre said:
that is the first transfer. i did a few drops of the syringe onto dish #1, then transfered it THAT dish: dish #2.
damn, atleast 4 sectors? that is crazy. i was reading someone's guide to doing agar ,i forget his name but it is his "thing", and my eye is still imperfect when it comes to sectoring. i guess i will aim for that think portion of myc between 3-6 o'clock.
frankhorrigan: what size dishes do you prefer? these are 50 or 60mmx15mm
can someone provide me with a reliable petri dish source? 100% sterile?
I'm going to you about the petris...
I use 100x15mm dishes, 3 section, similar to what RR uses in his videos.
If that is the first transfer, you have many more to do to achieve an isolate but....I recommend just inoculating grains that beautiful thing and cloning some fast-growing first flush fruits. Those will be some good fruiting isolates with less work and more reward than blind transfers.
My sterile procedures and agar write ups are in the works, I'm waiting to get my lightbox built because photographing petris is a pain in the ass. Soon though.
Is it stropharis' write up you are referring to?
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Stropharis



Registered: 04/02/07
Posts: 1,854
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: FrankHorrigan]
#18749581 - 08/23/13 11:22 PM (10 years, 5 months ago) |
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All projects down for several months at the moment, completing a move back to the good ol' US of A. Lots of work with the new abode, glad to help with the write-ups once I get back up. Seems all questions were answered here, no suprise with Frank around.
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FrankHorrigan
The Inquisition



Registered: 01/04/11
Posts: 10,573
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: Stropharis]
#18749632 - 08/23/13 11:38 PM (10 years, 5 months ago) |
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Oh there he is 
Good to see you around again!
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Stropharis



Registered: 04/02/07
Posts: 1,854
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Re: QUESTION ABOUT INOCULATING W/ AGAR (PICS) [Re: FrankHorrigan]
#18749800 - 08/24/13 12:35 AM (10 years, 5 months ago) |
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Yea Brother! If I'm not on the Shroomery, I am at the grindstone and trust me; I'd rather be at the Shroomery. New company, new place, new promotion, and new country. It will be about six months before I’m back at it, I did take a walk through a few large department stores though; it was like Disneyland for the mushroom cultivator. I saw a case of jars for like 15 dollars and I just stood there with my mouth open, and the tubs, wow! All over and dirt cheap!
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