Monday, March 19, 2007
Single stranded DNA cleavage
Mung Bean Nuclease ( 來自綠豆的精靈)
10X reaction buffer: (also for S1 nuclease)
300mM Na0Ac pH 5.0
500mM NaCl
1 or 10 mM ZnCl2 ( I tried both !)
50% Glycerol
37oC 15min (the reaction is highly efficient, 1u=1ug/min)
Any double-strand form is safe (dsDNA, DNA/RNA, dsRNA).
On the other hand, S1 can nick double strand DNA. S1 can also cleave
the "Nick" site to generate double-strand break.
10X reaction buffer: (also for S1 nuclease)
300mM Na0Ac pH 5.0
500mM NaCl
1 or 10 mM ZnCl2 ( I tried both !)
50% Glycerol
37oC 15min (the reaction is highly efficient, 1u=1ug/min)
Any double-strand form is safe (dsDNA, DNA/RNA, dsRNA).
On the other hand, S1 can nick double strand DNA. S1 can also cleave
the "Nick" site to generate double-strand break.