partial digestion - restriction conditions (Jan/17/2005 )
hi everyone
last time i asked for some tips about partial digestion, i forgot to give some details: i use 2 enzymes NcoI and HindIII, i cut pBluescript where my insert is put in. Generally I have 3 restriction sites in that insert: 2 for NcoI and 1 for HindIII.
What do You think about it?
How to manage that problem?
Thanks
Best
tomek
-tomgar-
Partial digestion is not a very good method to identify the recombinant clone. To this end, you may
1) reduce the digestion time;
2) reduce the restriction enzyme;
Also, you could choose other methods to complete the identification, such as sequencing, or restricting by other double=enzyme( one in the vector and another in the insert).
-mybbff-
QUOTE (mybbff @ Jan 19 2005, 08:33 PM)
Partial digestion is not a very good method to identify the recombinant clone. To this end, you may
1) reduce the digestion time;
2) reduce the restriction enzyme;
Also, you could choose other methods to complete the identification, such as sequencing, or restricting by other double=enzyme( one in the vector and another in the insert).
1) reduce the digestion time;
2) reduce the restriction enzyme;
Also, you could choose other methods to complete the identification, such as sequencing, or restricting by other double=enzyme( one in the vector and another in the insert).
i have to make partial digestion to obtain some DNA fragment needed to forward subcloning, it is necessary step for me. I will check both methods, thanks.
-tomgar-