sequencing problems

Yongin Kim yongland at netsgo.com
Thu May 13 05:07:33 EST 1999


Hello!

I am under genetic study revealing dinucleotide repeat alleles in human.
As a marker, I run sequenced puc18 vecter with my PCR samples at the same
time. I prepared the GATC with cycling sequencing protocol.
I see my result with silver staining which shows the yellow deposit on the
top of the sequencing gel after 1-2 hours running at the constant 55W
The GATC lanes all show identical single band with my PCR product(120-130
bp) on the top of the 6% denaturing gel.
I can't find the answer. I got always the same results.
Is this running problem or no cycling sequencing reaction
Is there anybody who encounder the same problem before?
Please let me know the way out ot this enigma.
Thanks for your suggestions





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