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Potvrzuji
  1. Úvodní strana
  2. Reagencie
  3. Life Technologies / Fermentas
  4. Conventional restriction enzymes
  5. BseJI (BsaBI)

BseJI (BsaBI)

Kód produktu: ER1711 Kód výrobce: ER1711 Kód dodavatele: {AFEFC569-2FBA-4059-9DB8-BF0CEEFFFB55} Výrobce: Life Technologies Czech Republic s.r.o.
BseJI (BsaBI)
2 686,20 Kč
2 220,00 Kč bez DPH
do týdne
2000 units

5'...G A T N N↓N N A T C...3'
3'...C T A N N↑N N T A G...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

  • Incubation at 37°C results in less than 10% activity.
  • Greater than 20-fold overdigestion with BseJI may result in star activity.
  • BseJI is blocked by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain, such as GM2163.

For methylation sensitivity refer to product specifications.

Conditions for 100% Activity
  • 1X Buffer O: 50 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 100 mM NaCl, and 0.1 mg/mL BSA
  • Incubate at 65°C
  • To ensure higher efficiency of digestion, perform the cleavage reaction under paraffin oil
Double Digestion Perform double digestion using DoubleDigest.
Hazardous No
Isoschizomers Search for commercial isoschizomers using REsearch.
Storage Buffer BseJI is supplied in:
10 mM Tris-HCl (pH 7.4 at 25°C), 100 mM NaCl, 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
O (Orange) 65°C NR 100† 100 NR NR 100† 2X†

† Star activity appears at a greater than 5-fold overdigestion (5 units x 1 hour).

Methylation Effects

  • Dam: may overlap - blocked.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - effect not determined.
Methylation type Sequence Cleavage effect
Dam (GATC) 5'...Gm6A TC(N)3ATC...3'
3'...C Tm6AG(N)3TAG...5'		 Blocked
CpG 5'...GAT(N)4ATm5C G...3'
3'...CTA(N)4TA Gm5C...5'		 No effect
CpG 5'...m5C GAT(N)4ATm5C G...3'
3'... Gm5CTA(N)4TA Gm5C...5'		 Not determined

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
21 2 2 1 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 1

 

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