unexpected band in PCR with plasmid
Posted 10 January 2013 - 04:24 AM
i have designed a set of primers for checking cloning in the plasmid pET28b. Though the primers are not expected to bring out any band with empty plasmid, it shows a band around 100bp which is also observed in the transformant plasmid. the primer sequences are:PET F- ATTCGGATCCACTAGTTATTG PET R- AATTCCCCTCTAGACCCTTGA. can anyone pls refer any online tool to get the sequence of the amplicons on submitting the template and primers.
Posted 10 January 2013 - 04:54 AM
Posted 10 January 2013 - 06:34 AM
Then if you do a primer blast (http://www.ncbi.nlm....s/primer-blast/) using just the perfect base paring parts of the primers, you get a product of 154bps.
Primer pair 1
Forward primer ATTCGGATCC
Reverse primer AATTCCCCTCTAGA
Product length 154
Posted 11 January 2013 - 01:07 AM