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Home | Molecular Biology | Restriction & Modifying Enzymes | Restriction Enzymes | Conventional

BclI

5'...T↓G A T C A...3'
3'...A C T A G↑T...5'
Available as a FastDigest enzyme for rapid DNA digestion

FastDigest enzyme available G buffer for 100% activity Optimal incubation at 55°C Star activity Cleavage blocked or impaired by Dam methylation Only small amounts (up to 10 u) can be inactivated at 80°C in 20 min LO certified

The BclI restriction enzyme recognizes T^GATCA sites and cuts best at 55°C in G buffer (Isoschizomers: FbaI, Ksp22I)
  
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Lambda DNA (dam-) digested with BclI

Lambda DNA (dam-) 0.7% agarose, 8 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

For methylation sensitivity refer to product specifications.

  
Storage Condition-20 C
HazardousNo
Compatible EndsBamHI, BglII, Bsp143I, MboI, PsuI.
Conditions for 100% Activity
  • Incubation at 37°C results in 50% activity
  • Greater than 15-fold over-digestion with BclI may result in star activity
  • Assayed using lambda DNA (dam-) (#SD0021)
  • BclI is blocked by dam methylation
  • To avoid dam methylation, use a dam-, dcm- strain, such as GM2163
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
Storage BufferBclI 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.

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
G (Green) 55°C 20-50 100 20-50 20-50 100† 100 1X† or 2X

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

Methylation Effects

Methylation type Sequence Cleavage effect
Dam(GATC)TGm6ATCA Blocked
EcoBI(TGA(N)8TGCT) 5'...TGm6A TCA(N)5 TGCT...3'
3'...AC Tm6AGT(N)5m6ACGA...5'		 Blocked

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
8 0 0 0 0 0
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
0 0 0 0 0 1

New sites generated by ligation

Newly generated recognition sites resulting from ligation of protruding compatible DNA ends

Recognition Sequence Second Enzyme Enzymes recognizing newly generated recognition sequence
T^GATCA
  • BamHI/FastDigest BamHI (G^GATCC)
  • BglII/FastDigest BglII (A^GATCT)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I) (^GATC)
  • MboI/FastDigest MboI (^GATC)
  • PsuI (BstYI)/FastDigest PsuI* (R^GATCC)
  • PsuI (BstYI)/FastDigest PsuI* (R^GATCT)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI

Newly generated recognition sites resulting from fill-in of 5'-overhang and self-ligation

Recognition Sequence Newly generated sequence after reaction Enzymes that cleave the newly generated sequence
T^GATCATGATCGATCA
  • [Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)]
  • Bsu15I (ClaI)/FastDigest ClaI (Bsu15I)
  • [DpnI/FastDigest DpnI]
  • [MboI/FastDigest MboI]
  • TaqI/FastDigest TaqI