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OfflineHafrican
Mega-ultrachicken

Registered: 01/31/03
Posts: 105
Loc: SW Azerbaijan
Last seen: 17 years, 11 days
Slow growth on peroxide agar -- HELP?
    #1633432 - 06/14/03 04:09 PM (21 years, 1 month ago)

So, Jim has been cloning on peroxide agar for awhile. He has experienced extremely slow growth. All of the cloning was first generation (i.e. from mushrooms germinated from spores). He is using premixed MEA in the ratio of 7g MEA to 150 mL water. He is sterilizing the mix at 15 psi for 20 minutes. Jim is a lazy bitch, and tends to let his plates sit around (wrapped in cling wrap) for a few days -- no visible contamination occurs during this time. After the tissue transfer, he wraps the edges in parafilm and incubates at around 80-86 degrees. The tissue samples are about 1/8" square. Any explanation? What are your friends average times for 3/4 to full colonization of a (peroxide) agar plate? Jim is frustrated and is going to go back to liquid culture ....

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Invisiblemicro
bunbun has a gungun
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Registered: 05/09/03
Posts: 7,532
Loc: Brick City
Re: Slow growth on peroxide agar -- HELP? [Re: Hafrican]
    #1633484 - 06/14/03 04:35 PM (21 years, 1 month ago)

Mycelium usually grows more slowly on peroxidazed agar, but you should only use parafilm when storing in the fridge, not when incubating. Just keep them in a semi-sterile enviornment w/ no parafilm when incubating; you can even stack them and put them back into the original sleeve (which should be sterile.) Make sure it's free from drafts, etc.

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Micro


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OfflineHafrican
Mega-ultrachicken

Registered: 01/31/03
Posts: 105
Loc: SW Azerbaijan
Last seen: 17 years, 11 days
Re: Slow growth on peroxide agar -- HELP? [Re: micro]
    #1633796 - 06/14/03 08:04 PM (21 years, 1 month ago)

Can Jim use cling wrap on the edges while incubating? He is concerned about crap getting in through the sides of his glass dishes. Also, when he incubates in a bag (as suggested), it condensates something awful (the plates and bags). Thanks.

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Invisiblemicro
bunbun has a gungun
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Registered: 05/09/03
Posts: 7,532
Loc: Brick City
Re: Slow growth on peroxide agar -- HELP? [Re: Hafrican]
    #1633817 - 06/14/03 08:11 PM (21 years, 1 month ago)

It is not reccomended (by scientists) that you wrap them while incubating, but you can put them back in the sleeve like I stated above.

To prevent condensation you can flip the petrie dishes upside down once visible germination is present; it does not inhibit growth.

--
Micro


--------------------
Any research paper or book for free
(Avatar is Maxxy, a character by Mizzyam, RIP)

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Invisible40oz
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Registered: 01/18/01
Posts: 30,119
Loc: Sandy Eggo. Ca.
Re: Slow growth on peroxide agar -- HELP? [Re: Hafrican]
    #1634492 - 06/15/03 02:20 AM (21 years, 1 month ago)

how many cc's of h202 are you using per ____ml flask?


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