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Potvrzuji
  1. Úvodní strana
  2. Reagencie
  3. Life Technologies / Fermentas
  4. Conventional restriction enzymes
  5. BplI

BplI

Kód produktu: ER1311 Kód výrobce: ER1311 Kód dodavatele: {114B9425-EB61-4B72-B05E-BA0A95064285} Výrobce: Life Technologies Czech Republic s.r.o.
BplI
2 071,52 Kč
1 712,00 Kč bez DPH
do týdne
100 Units

5'...↓ 8(N) G A G (N)5 C T C (N)13↓...3'
3'...↓13(N) C T C (N)5 G A G (N)8 ↓...5'

Zobraz detailní popis

Detailní popis

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

BplI requires only Mg2+ for its activity, but is stimulated by S-adenosylmethionine. 0.05 mM S-adenosylmethionine gives more than a  100-fold increase in BplI activity. Still, complete cleavage of some substrates with BplI is difficult to achieve. BplI concentration is determined by the maximum cleavage level achieved when no change in the fragmentation pattern is observed with further enzyme increase.

For methylation sensitivity refer to product specifications.

Conditions for 100% Activity
  • [1x Buffer Tango] + SAM: [33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, and 0.1 mg/mL BSA] + 0.05 mM S-adenosylmethionine
  • Incubate at 37°C
Digestion of Agarose-embedded DNA Minimum 10 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double Digestion Perform double digestion using DoubleDigest.
Hazardous No
Isoschizomers Search for commercial isoschizomers using REsearch.
Storage Buffer BplI is supplied in: 10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM KCl, 1 mM DTT, 1 mM EDTA, 0.2 mg/mL BSA, and 50% (v/v) glycerol.
Storage Condition -20 C

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 0-20 (+SAM) 20-50 (+SAM) 0-20 (+SAM) 0-20 (+SAM) 100 (+SAM) 20-50 (+SAM) 1X (+SAM)

Methylation Effects

  • Dam: never overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - no effect.
  • EcoKI: may overlap - no effect.
  • EcoKI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
CpG 5'...m5C GAG(N)5CTm5C G...3'
3'... Gm5CTC(N)5GA Gm5C...5'		 No effect

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
1 0 0 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 0

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