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MbiI (BsrBI)

5'...C C G↓C T C...3'
3'...G G C↑G A 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 CpG methylation Thermal inactivation at 65°C in 20 min Genome qualified LO certified

The MbiI (BsrBI) restriction enzyme recognizes CCGCTC(-3/-3)^ sites and cuts best at 37°C in Tango buffer (Isoschizomers: AccBSI, BsrBI)
  
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Lambda DNA digested with MbiI (BsrBI)

Lambda DNA digested with MbiI (BsrBI), 1.4% agarose, 17 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
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
Digestion of Agarose-embedded DNAMinimum 5 units of the enzyme are required for complete digestion of 1 µg of agarose-embedded lambda DNA in 16 hours.
Double DigestionPerform double digestion using DoubleDigest.
IsoschizomersSearch for commercial isoschizomers using REsearch.
NoteLow salt, high glycerol (> 5%) concentrations, pH > 8.0 or a large excess of enzyme may result in star activity.
Storage BufferMbiI 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 20-50 100 0-20 20-50 100 20-50 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
CpGGAGCGG Impaired
CpG 5'...m5C GAGCGG...3'
3'... Gm5CTCGCC...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 0-20 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
17 1 4 2 3 3
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
4 4 5 5 3 2

New sites generated by ligation

Newly generated recognition sites resulting from ligation of blunt DNA ends

Recognition sequence Second restriction enzyme Restriction enzymes cleaving the newly generated recognition sequence
CCG^CTC
  • AjiI (BmgBI)* (CAC^GTC)
  • AjiI (BmgBI)* (GAC^GTG)
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I) (TAC^GTA)
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I) (CAC^GTG)
  • FspAI/FastDigest FspAI* (RTGC^GCAC)
  • FspAI/FastDigest FspAI* (RTGC^GCAT)
  • HincII (HindII)/FastDigest HincII* (GTY^GAC)
  • NsbI (FspI)/FastDigest FspI (NsbI) (TGC^GCA)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTA)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTG)
  • ZraI (GAC^GTC)
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • AluI/FastDigest AluI (AG^CT)
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • CviJI* (RG^CT)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • HpyCH4V (TG^CA)
  • MlsI (MscI)/FastDigest MscI (MlsI) (TGG^CCA)
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • Bsh1236I (BstUI)/FastDigest Bsh1236I (CG^CG)
  • Bsp68I (NruI)/FastDigest NruI (RruI) (TCG^CGA)
  • Bsh1236I (BstUI)/FastDigest Bsh1236I
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • Bst1107I (BstZ17I)/FastDigest BstZ17I (Bst1107I) (GTA^TAC)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • MbiI (BsrBI)/FastDigest BsrBI (MbiI)
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • CviJI
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • MspA1I* (CMG^CGG)
  • BseDI (BsaJI)/FastDigest BsaJI (BseDI)
  • Bsh1236I (BstUI)/FastDigest Bsh1236I
  • BtgI
  • Cfr42I (SacII)
  • MspA1I
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • MspA1I* (CMG^CTG)
  • PvuII/FastDigest PvuII (CAG^CTG)
  • MspA1I
  • SsiI (AciI)/FastDigest AciI (SsiI)
  • PdiI (NaeI)/FastDigest NaeI (PdiI) (GCC^GGC)
  • BcnI (NciI)/FastDigest NciI (BcnI)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BssKI
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
  • SmaI/FastDigest SmaI (CCC^GGG)
  • SrfI (GCCC^GGGC)
  • BcnI (NciI)/FastDigest NciI (BcnI)
  • Bme1390I (ScrFI)/FastDigest ScrFI (Bme1390I)
  • BseDI (BsaJI)/FastDigest BsaJI (BseDI)
  • BssKI
  • HpaII/FastDigest HpaII
  • MspI (HpaII)/FastDigest MspI
GAG^CGG
  • AjiI (BmgBI)* (CAC^GTC)
  • AjiI (BmgBI)* (GAC^GTG)
  • Eco105I (SnaBI)/FastDigest SnaBI (Eco105I) (TAC^GTA)
  • Eco72I (PmlI)/FastDigest PmlI (Eco72I) (CAC^GTG)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTA)
  • Ppu21I (BsaAI)/FastDigest BsaAI (Ppu21I)* (YAC^GTG)
  • ZraI (GAC^GTC)
  • SetI
  • AluI/FastDigest AluI (AG^CT)
  • CviJI* (RG^CT)
  • MspA1I* (CMG^CTG)
  • PvuII/FastDigest PvuII (CAG^CTG)
  • AluI/FastDigest AluI
  • CviJI
  • SetI
  • Bst1107I (BstZ17I)/FastDigest BstZ17I (Bst1107I) (GTA^TAC)
  • Csp6I (CviQI)/FastDigest Csp6I
  • RsaI/FastDigest RsaI
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI) (GG^CC)
  • CviJI* (RG^CC)
  • Eco147I (StuI)/FastDigest StuI (Eco147I) (AGG^CCT)
  • Eco47III (AfeI)/FastDigest AfeI (Eco47III) (AGC^GCT)
  • MlsI (MscI)/FastDigest MscI (MlsI) (TGG^CCA)
  • CviJI
  • DpnI/FastDigest DpnI (GA^TC)
  • HinfI/FastDigest HinfI
  • PleI
  • SchI (MlyI)/FastDigest MlyI (SchI)
  • Ecl136II (EcoICRI)/FastDigest Ecl136II (GAG^CTC)
  • AluI/FastDigest AluI
  • Alw21I (BsiHKAI)/FastDigest Alw21I
  • CviJI
  • Ecl136II (EcoICRI)/FastDigest Ecl136II
  • Eco24I (BanII)
  • SacI/FastDigest SacI
  • SduI (Bsp1286I)/FastDigest Bsp1286I (SduI)
  • SetI
  • Eco32I (EcoRV)/FastDigest EcoRV (Eco32I) (GAT^ATC)
  • Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)
  • DpnI/FastDigest DpnI
  • MboI/FastDigest MboI
  • EheI (SfoI)/FastDigest EheI (GGC^GCC)
  • BsuRI (HaeIII)/FastDigest HaeIII (BsuRI)
  • CviJI