RNase protection using PCR products
salger at wap1.zi.biologie.uni-muenchen.de
Mon Aug 7 17:12:16 EST 1995
HOFSTRA ANGELA 905-822-3520 (Hofstra at aa.wl.com) wrote:
: I'm interested in using PCR products for making riboprobes for RNase protection
: I have a technical bulletin from Ambion that give a brief description; but, if
: anyone has a reliable method for introducing a relevant promoter site and
: subsequent transcription I'd love to read about it. Please respond or post.
: Thanks, Angela
you just need an oligo which contains the T7 promoter at the 5' end and some
specific, complementary bases.
Here's what my primers looked like:
5' T7 promoter 3'
gtaatacgactcactataggg-20bp of complementary sequence of the 3'-end of my gene
20bp of complementary sequence of the 5'-end
-> PCR -> in vitro transcription
Please tell me, if you need more specific informations.
Klaus Salger phone : +49 (0)89 5902 -502
Zoologisches Institut FAX : -450
AG MacWilliams e-mail: salger at zi.biologie.uni-muenchen.de
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