300 bp more after PCR purification - please help - (Jan/17/2008 )
I got pretty nice PCR products (cytochrom b, DNA from tissue). Bur after cleaning them with PCR purification kit GENOMED a few of samples "jumped from the row" and are 300 to 400 bp longer (picture of one example attached). Eluted into TE buffer. Shall I proceed to sequencing? I would highly appreciated any comment, many thanks, Lubos
-lubos-
QUOTE (lubos @ Jan 17 2008, 07:03 PM)
I got pretty nice PCR products (cytochrom b, DNA from tissue). Bur after cleaning them with PCR purification kit GENOMED a few of samples "jumped from the row" and are 300 to 400 bp longer (picture of one example attached). Eluted into TE buffer. Shall I proceed to sequencing? I would highly appreciated any comment, many thanks, Lubos
When in doubt, I sequence. It's inexpensive and extremely helpful. Your result is unusual -- sequence away.
-Cheamps-
i would sequence them.
-scolix-
I vote for sequencing too.
As for your observation if you have salt in your DNA mix, it tends to retard the movements of said DNA band...though I have not seen such a clear band shift before. Protein binding to your DNA can also effect a retardation.... Still this is pretty odd.
Sequence it.
-perneseblue-