Cloning in pCR2.1 plasmid
Posted 09 August 2010 - 05:54 AM
Posted 10 August 2010 - 05:10 PM
If you used genomic DNA, did you blast your primers against the genomic data base to see if the primers bind any other site aside from your gene of interest?
Posted 12 August 2010 - 08:15 AM
Posted 14 August 2010 - 07:05 PM
DNA, i am assuming it is genomic DNA?
DId you make the cDNA?
Did you do a blast search on the primers to make sure that there are no secondary binding sites?
What is the tm of your primers?
Posted 15 August 2010 - 03:41 PM
Posted 15 August 2010 - 04:08 PM
How exactly did you do this?
I'm sure you used the right primers, however, your assumption that nothing else could have been amplified is likely incorrect.
If you have a sequence that includes both vector and insert, you should be able to easily identify your insert sequence and isolate it for BLAST analysis. What is it you've cloned?
Posted 16 August 2010 - 01:02 PM
Posted 16 August 2010 - 01:08 PM