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Hin1I (BsaHI)

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

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

The Hin1I (BsaHI) restriction enzyme recognizes GR^CGYC sites and cuts best at 37°C in G buffer (Isoschizomers: AcyI, BsaHI, BssNI, BstACI, Hsp92I)
  
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Lambda DNA digested with Hin1I (BsaHI)

Lambda DNA digested with Hin1I (BsaHI), 1% agarose, 40 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

Hin1I 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
Compatible EndsBsp119I, Bsu15I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, SsiI, TaqI, XmiI.
Conditions for 100% Activity
  • 1X Buffer G: 10 mM Tris-HCl (pH 7.5 at 37°C), 10 mM MgCl2, 50 mM NaCl, 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.
Storage BufferHin1I is supplied in: 10 mM potassium phosphate (pH 7.4 at 25°C), 100 mM NaCl, 1 mM EDTA, 1 mM DTT, 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) 37°C 20-50 100 20-50 20-50 20-50 20-50 1X or 2X

Methylation Effects

Methylation type Sequence Cleavage effect
Dcm (CCWGG) 5'...GRCGCm5CW GG...3'
3'...CYGCG GWm5CC...5'		 Impaired
CpGGRCGYC 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
40 7 1 6 3 3
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
1 1 1 1 2 4

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
GA^CGYC
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I) (G^CGC)
  • SsiI (AciI)/FastDigest AciI (SsiI)* (C^CGC)
  • CseI (HgaI)/FastDigest HgaI (CseI)
  • MaeII (A^CGT)
  • Psp1406I (AclI)/FastDigest AclI (Psp1406I) (AA^CGTT)
  • MaeII
  • SetI
  • TaiI (MaeII)/FastDigest TaiI
  • NarI (GG^CGCC)
  • CseI (HgaI)/FastDigest HgaI (CseI)
  • Hin1I (BsaHI)/FastDigest BsaHI (Hin1I)
GG^CGYC
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I) (G^CGC)
  • SsiI (AciI)/FastDigest AciI (SsiI)* (C^CGC)
  • HhaI/FastDigest HhaI
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • NarI (GG^CGCC)
  • BbeI
  • FastDigest HaeII (BfoI)
  • BshNI (BanI)/FastDigest BanI (BshNI)
  • BspLI (NlaIV)/FastDigest NlaIV (BspLI)
  • EheI (SfoI)/FastDigest EheI
  • HhaI/FastDigest HhaI
  • Hin1I (BsaHI)/FastDigest BsaHI (Hin1I)
  • Hin6I (HinP1I)/FastDigest HinP1I (Hin6I)
  • NarI
  • SspDI (KasI)

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
GR^CGYCGRCGCGYC
  • Bsh1236I (BstUI)/FastDigest Bsh1236I