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Offlinemesq
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Registered: 02/24/02
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Streaking technique and Plastic inoculation loops
    #1448522 - 04/11/03 01:51 AM (17 years, 5 months ago)

I just have a question regarding the above... In alot of streaking techniques I see that you must flame the loop after streaking each quadrant of the plate.. so as to properly dilute the spores for isolating cultures... Since I have meltable inoculation loops should I use a new loop for each quadrant or just use the same one??

Does anyone know what I mean ?


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InvisibleJoshua
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Registered: 10/27/98
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Re: Streaking technique and Plastic inoculation loops [Re: mesq]
    #1448811 - 04/11/03 05:31 AM (17 years, 5 months ago)

I would either rinse the loop in alcohol or use a new one.

Joshua


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Invisiblewoodrow
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Registered: 03/17/03
Posts: 142
Re: Streaking technique and Plastic inoculation loops [Re: mesq]
    #1448838 - 04/11/03 06:19 AM (17 years, 5 months ago)

Use one loop and streak the entire plate. Quadrant streaking is for bacteria where you need to dilute a large population of organisms. Shroom spores tend to stick together in large clumps and the problem when using an innoculating loop is more often too few colonies rather than too many. I like to put a drop or two of spore suspension on the plate and let it roll around. No streaking necessary.
When colonies do form, they are usually the result of the germination of a large number of spores from a single clump. Shrooms need the combined genetic information from several individual spores if they are to form fruiting bodies and the gene pool is distributed over a wide area of the mycelium. When you pick colonies from the plate you need to scoop up at least a quarter inch of mycelium to get the required gene pool. Touching the colony with a loop, the way you do with bacteria, to make a transfer should not be done with shrooms.


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Offlinemesq
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Re: Streaking technique and Plastic inoculation loops [Re: woodrow]
    #1448847 - 04/11/03 06:28 AM (17 years, 5 months ago)

TThe print that I am working on is definitely unsterile hence why I am using this kind of isolation technique... so I can dilute the different spores. Is this the wrong way to isolate a culture from an unsterile source?


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Invisiblewoodrow
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Registered: 03/17/03
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Re: Streaking technique and Plastic inoculation loops [Re: mesq]
    #1451694 - 04/11/03 09:00 PM (17 years, 5 months ago)

Your streaking tek is good but shroom spores don't streak well because of their chunkiness so you are likely to get spore germination near the inoculation site and nothing but contams on the rest of the plate. The important thing is to get spore germination and see what kind of contams you have and how they grow so you can customize your methods for the next isolation. If your contams are yeasts or bacteria that form nice firm colonies, you can transfer a piece of mycelium to the center or edge of another agar plate- but don't streak. Let the mycelium grow onto the empty area of the plate, leaving the contams behind, and pick another pice of mycelium from the advancing edge of growth for a clean isolation onto a third plate.
If the contams are bacteria, peroxide or peroxide agar are often helpful. The worst contams are molds and swarming bacteria. With these you need to make rapid transfers and keep on streaking. An extra solid agar helps to prevent swarming. Molds are a problem because shroom mycelium starts out looking exactly like mold mycelium and peroxide is useless against molds. Once a mold begins to disperse spores, the whole plate is toast.
Good luck!


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InvisibleJoshua
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Re: Streaking technique and Plastic inoculation loops [Re: woodrow]
    #1451795 - 04/12/03 06:04 AM (17 years, 5 months ago)

Good point...I missed that one.

Joshua


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Great books for inquiring minds!

"Life After Death is Saprophytic!"


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Offlineshirley knott
not my real name
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Registered: 11/11/02
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Re: Streaking technique and Plastic inoculation loops [Re: woodrow]
    #1451853 - 04/12/03 07:44 AM (17 years, 5 months ago)

woodrow, i've read a few of your posts now, and you certainly know your shit. respect due - i'll search by your name when i want good agar advice, for one.

have some shrooms :grin: 


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buh


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Invisibleno-tone
Enema Bag Jones

Registered: 10/17/00
Posts: 1,091
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Re: Streaking technique and Plastic inoculation loops [Re: shirley knott]
    #1452102 - 04/12/03 12:27 PM (17 years, 5 months ago)

I feel the same way. More shrooms for woodrow!


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Man thinks. God laughs. - Jewish Proverb


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