Thermo Scientific™ BspLI (NlaIV)
Cut at GGN^NCC sites with BspLI (NlaIV) restriction enzyme, which performs best at 37°C in Tango buffer (Isoschizomers: BmiI, NlaIV, PspN4I).
Manufacturer: Thermo Scientific™ ER1151
Conventional restriction endonucleases are a large collection of high quality restriction enzymes, optimized to work in one of the buffers of the Five Buffer System. In addition, the universal Tango buffer is provided for convenience in double digestions. All of the enzymes exhibit 100% activity in the recommended buffer and reaction conditions. To ensure consistent performance, Thermo Scientific restriction enzyme reaction buffers contain premixed BSA, which enhances the stability of many enzymes and binds contaminants that may be present in DNA preparations.
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).