Cut at GGN^NCC sites with BspLI (NlaIV) restriction enzyme, which performs best at 37°C in Tango buffer (Isoschizomers: BmiI, NlaIV, PspN4I).
Brand: Thermo Scientific ER1152
UNSPSC : 12352204
Code : 88
Additional Details : Weight : 0.00900kg
5'..G G N▵N C C...3'
3'..C C N▵N G G...5'
Conditions for 100% Activity:
- 1X Buffer Tango: 33mM Tris-acetate (pH 7.9 at 37°C), 10mM Mg-acetate, 66mM K-acetate and 0.1mg/mL BSA
- Incubate at 37°C
- BspLI is supplied in: 10mM Tris-HCl (pH 7.4 at 25°C), 200mM NaCl, 1mM DTT, 1mM EDTA, 0.2mg/mL BSA and 50% (v/v) glycerol
Ligation and Recleavage:
- After 50-fold overdigestion with BspLI, approximately 90% of the DNA fragments can be ligated and more than 95% of these can be recut
- Dam: never overlaps — no effect
- Dcm: may overlap — cleavage impaired
- CpG: may overlap — cleavage impaired
- EcoKI: never overlaps — no effect
- EcoBI: never overlaps — no effect
BspLI cleavage is impaired by overlappingdcm methylation. To avoiddcm methylation, use adam-, dcm strain such as GM2163 (#M0099).