In article <53h25p$1rp at is.bbsrc.ac.uk> Clive Tregaskes,
Clive.tregaskes at bbsrc.ac.uk writes:
>1 two start points (tho' Northern suggests this isn't so)
>2 short transcripts are due to secondary structure stopping the RT
>3 long transcript is due to DNA contamination of my RNA prep (but
>contains a dC tail)
>Lots of ideas but no answers, so does anybody know how you tell the
>real start site from any secondary structure artifacts.
Design a primer to the 5' end of the new sequence and do standard rtPCR
to see if you get the predicted fragment.
Use the novel sequence as a probe on a northern blot and see if you pick
up the predicted message.
Use the new sequence on a genomic southern and see if it maps to the same
fragment as the rest of the sequence (need to maybe use a rare cutter)
Hope this helps.
**my opinions are just that**
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Eric Lader Ph.D. Senior R&D Scientist,
Ambion Inc.
Unique tools for Molecular Biology Research
Phone (800) 888-8804 Fax (512) 445-7139
(512) 445-6979 lader at ambion.com
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