Dear all,

A couple of months ago I sent an email reporting rather frustrating 
sequencing problems. It appears we have found the source: The UV light 
box for cutting bands!

When we narrowed down the problem to the gel cutting stage we proceeded 
by sequencing directly from a purified PCR product (even though we 
often get unspecific bands). The sequences were mostly ok, of course 
this protocol was no good for cloning. During our further 
investigations we found out that someone changed the light bulb around 
the time we had the first problems. Although the bulb hasn't been 
replaced yet it appears that cutting bands from Ethidium bromide gels 
is more productive than a cheap cyber green stain that we have started 
using in February. We also keep the UV exposure to an absolute minimum!

Anyway, thank you all for the many suggestions. And to the couple of 
people who reported similar problems: Go and check you UV light bulb!

Best wishes, Birgit

Birgit Meldal, PhD
Division of Transfusion Medicine
Department of Haematology
University of Cambridge
National Blood Service Center
Long Road
Cambridge
CB2 2PT

Tel: +44 (0) 1223 548049
E-mail: bhmm2@cam.ac.uk

bhmm2@cam.ac.uk