We have some problems with blue dye blobs around 100-150 bp when
we use the Perkin Elmer recommeded protocol to remove residual
BigDye terminators. We precipitate in microamp trays using either
60% isopropanol or 60% ethanol. The problem is more pronounced
when the sequencing template is a short PCR product. We never see
the blobs when we clean up with the p-30 Micro Bio-spin columns
from Bio-Rad. Unfortunately this product is not available in a 96
well format. We have previously used the 96 well gel filtration
kit from AGTC but the plates do not work properly for BigDye
terminators.Any suggestions?
Best regards,
Peter Bjarke Olsen
Bioinformatics and DNA Sequencing
Novo Nordisk A/S
