Hello.
I have some human ss cDNA from primer extension and was wondering what the best
method was to sequence it.
There do not appear to be any vectors which it can be inserted into, as they
all require dsDNA.
That leaves me to sequence it manually with Sequenase v2.0 or equivalent. My
main questions are:
What should I use as a template?
Should I do an RT using oligo(dT) primer, or should I use my specific primer
(from the p. extn) for the template? My main concern is the orientation of
the strand. My 5'-3' mRNAs and my 5'-3' primer in the same orientation. Hmmm.
Please help.
Andrea.
-----------------------------------------------
Andrea Varga
The Department of Surgery
The Queen Elizabeth Hospital
WOODVILLE SA 5011
Telephone (W): (08) 8222 7355
Fax (W): (08) 8222 6028
Mobile: 0414 604 258
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