size of gene insert wrong? - (Mar/02/2009 )
I performed a double digestion using BamH1 and HindIII on a strain of E. Coli. accorrding to the protein sequnce, the size of the insert should be 82 bp. But after the double digestion and looking at it on my agarose gel, I calculated that the insert was 0.745 kb. This is a really big difference....what went wrong here?
-eunicycle-
eunicycle on Mar 2 2009, 05:14 PM said:
I performed a double digestion using BamH1 and HindIII on a strain of E. Coli. accorrding to the protein sequnce, the size of the insert should be 82 bp. But after the double digestion and looking at it on my agarose gel, I calculated that the insert was 0.745 kb. This is a really big difference....what went wrong here?
Hi
your RE have other restriction sites, not unique to your sequence.
-dormancy-
82 bp is kind of too small to see on agaros gel after double digestion.
-pcrman-
pcrman on Mar 2 2009, 09:42 PM said:
82 bp is kind of too small to see on agaros gel after double digestion.
Yea i have to agree with pcrman. and perhaps u should double check on the sequence of the DNA
-hanming86-