PCR and the Central Dogma?

Petur Henry Petersen php at rhi.hi.is
Mon Jul 19 09:12:40 EST 1993


The central dogma, fraud or what?

Doing a routine work of sequencing a 300 bp region of the cytochrome b
gene in fish mtDNA, (amplified by PCR, a la Kocher et al. 1989, in
PNAS) I suddenly got puzzling results. I use two universal primers
L14841 and H15149, amplify a single stranded DNA with the L-primer and
I use the H-primer for sequencing. The DNA sequences I got from two
different species at the same time was as follows, (imediately starting
from the H-primer):

5'end-L-primer(3'->5')-tg-H*-primer(3'->5')-5'-245 bp of the cyt b gene-3'end.

What is unusual I got the L primer and the template (H*) for H primer in the 
beginning of my sequence, but more puzzling is the direction they were
sequenced, e.g. from the 3' end to the 5' end. 

It seem to me that these results are contradicting what I have learned,
so I would like to get comment on this.

Snaebjorn Palsson
University of Iceland
sp at lif.hi.is



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