Restriction problemsrestriction plasmid Bin
Posted 08 October 2012 - 07:14 AM
I have been trying to clone my insert into binary vector. However after trying for 15 days I find that the problem is restriction of plasmid. I am unable to restrict the plasmid and have also tried different restriction sites. I have used this plasmid earlier and it worked fine. I have tried restricting the entire cassette from the plasmid and I find that there is partial restriction.
The plasmid is ~16kb binary vector and I have cloned it in Dh5 alpha cells.
Has anyone face this kind of problem. If so please help me out.
Posted 08 October 2012 - 09:35 AM
Posted 08 October 2012 - 10:03 AM
I do multiple mini prep to get the plasmid (16kb binary vector). I wonder if this is contributing to inhibitors. Should I do a midi prep.
Posted 08 October 2012 - 10:08 AM
Posted 08 October 2012 - 10:38 AM