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production of antisense oligonucleotide by PCR ?

rogier stugerNOstSPAM at cellbiology.uni-frankfurt.de.invalid
Thu Jan 20 13:54:54 EST 2000


Hiya,
the tricky part is separating the strands, then isolating the strand
you need.
A  much much much easier method is to make riboprobes.
Just PCR your 200 bp piece of DNA with primers that contain T7 and T3
promoters as overhangs.
By in vitro transcription with T7 or T3 RNA polymerase, you can make
sense or antisense probes. Simply label them by adding
radioactive/fluorescent/DIGged/biotinylated/etc. rNTPs to your
transcription mix.
Good luck,
Rogier
Dept MicFizz
Free U of A
Amsterdam, The Netherlans


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