is there any way to "hide" one restriction site of two in order to dig - (Jul/31/2006 )
i'd like to move a two gene construct across to another plasmid; unfortunately, the enzyme i'd like to use at the 3' end is present within the construct. is there any way to silence the one site while still leaving the other viable? my other alternative is to use an enzyme at the 3' end that will add an extra 1kb to my insert.
thx in advance.
you may do partial digestion on your construct using limited amouts of RE, gel purify the band with right length, and then clone it.
I would rather pcr amplify and then clone.
To begin with, I would try partial digest and try ligation.
If this doesnt work, then PCR amplify with different sites.