DNA Isolation Protocol
DELANEYT at am.abru.cg.com
Tue Apr 4 13:32:06 EST 1995
Lynn Reuber asks if..
>anyone out there have a good protocol for making plant DNA
>from 1-2 g of tissue without dealing with CsCl? Has
>anyone had experience with making DNA from seedlings?
Lynn, ..I tried to reply to you directly, but my mailer won't reply to your
address, and you didn't provide your e-mail address, ...so at the risk of
boring others, my reply is shown below.
I'll take this chance to request that persons posting messages to the BB
should include their e-mail address in the body of their message ;-)
I have had good success with the procedure in Dellaporta et al, Plant Molec
Biol Reporter 1: 19-21. The protocol also works well when scaled down to an
eppindorf-size prep on 15-500 mg tissue, such as for small seedlings (ie
scale down reagents to 600 ul Ex Buf, 40 ul SDS, 200 ul KOAc, 400 ul
isopropanol). For RE digestion adding spermidine to 5 mM often makes a big
difference (add AFTER combining DNA and RE buffer/salts to prevent
spermidine pptn of DNA!).
Ciba Ag Biotech
delaneyt at am.abru.cg.com
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