Base pairs apart between 2 sites for sucessful double digestion? - (Jan/18/2007 )
How many base pairs apart between 2 restriction sites are sufficient for sucessful double digestion? If there are 12 bp apart for NheI and XbalI, is it OK? Thanks!
-plant-
12 bps is more than sufficient.
-scolix-
12 bp is sufficent in your exp.
For such purpose, you may have a look at neb webpage "cleavage close to DNA ends". Some enzymes need more than 6 bases some are ok with just2. So that may be helpful for sequential digestion.
-fred_33-