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BspLI (NlaIV)

5'...G G N↓N C C...3'
3'...C C N↑N G G...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available Tango buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by overlapping Dcm methylation Cleavage blocked or impaired by overlapping CpG methylation Thermal inactivation at 65°C in 20 min LO certified

The BspLI (NlaIV) restriction enzyme recognizes GGN^NCC sites and cuts best at 37°C in Tango buffer (Isoschizomers: BmiI, NlaIV, PspN4I)
  
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Lambda DNA digested with BspLI (NlaIV)

Lambda DNA digested with BspLI (NlaIV), 1.4% agarose, 82 cleavage sites

Thermo Scientific 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.

Highlights

  • Superior quality – stringent quality control and industry leading manufacturing process
  • Convenient color-coded Five Buffer System
  • Includes universal Tango buffer for double-digestions
  • BSA premixed in reaction buffers
  • Wide selection of restriction endonuclease specificities

Applications

  • Molecular cloning
  • Restriction site mapping
  • Genotyping
  • Southern Blot
  • Restriction fragment length polymorphism (RFLP)
  • SNP

Note

BspLI cleavage is impaired by overlapping dcm methylation. To avoid dcm methylation, use a dam-, dcm- strain, such as GM2163.

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Conditions for 100% Activity
  • 1X Buffer Tango: 33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate,and 0.1 mg/mL BSA
  • Incubate at 37°C
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferBspLI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 200 mM NaCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/ml BSA, and 50% (v/v) glycerol.

Reaction conditions

Recommended buffer for 100% activity Optimal temperature Enzyme activity in Thermo Scientific buffers, % Tango buffer for double digestion
B (blue)
1X		 G (green)
1X		 O (orange)
1X		 R (red)
1X		 Tango
(yellow)
1X / 2X
Tango 37°C 50-100 50-100 0-20 20-50 100 20-50 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
Dcm (CCWGG) 5'...GGNNCm5CW GG...3'
3'...CCNNG GWm5CC...5'		 No effect
Dcm (CCWGG) 5'...Cm5CW GGNNCm5CW GG...3'
3'...G GWm5CCNNG GWm5CC...5'		 Blocked
CpG 5'...GGNNCm5C G...3'
3'...CCNNG Gm5C...5'		 No effect
CpG 5'...m5C GGNNCm5C G...3'
3'... Gm5CCNNG Gm5C...5'		 Blocked

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
82 6 18 24 11 12
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
13 14 15 15 10 23