Digest with BglII and SalI - (Aug/07/2008 )
I prepare to digest my plasmid pEGFP-C1 with BglII and SalI. Can I use these enzymes at the same time or one by one. If I use one by one, which enzyme used first is better. And how can I check wether my plasmid is digest with both enzyme (just about 24bp between 2 enzyme)
Looking forward your suggestion
if you have purchase the new NEB SalI, technically you can use both BglII and SalI together is a simultaneous digest.
If you use the enzyme sequentially, start with SalI first, followed by BglII. BglII is a more robust enzyme
You can't. That is why the least efficient enzyme and the enzyme that required the most bp skirting it restriction site is used first, thus allowing you to see if the digest has worked.
However a strong warning of caution, many people have experienced difficulties in their ligation when SalI is used. Look up NEB website, SalI has a few problem with it. Is there any way to avoid using SalI? XhoI form compatible ends with SalI