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Potvrzuji
  1. Úvodní strana
  2. Reagencie
  3. Life Technologies / Fermentas
  4. Conventional restriction enzymes
  5. Bsp143I (Sau3AI)

Bsp143I (Sau3AI)

Kód produktu: ER0782 Kód dodavatele: {28CAC371-9584-4FAF-BAD0-A09E982A734E} Výrobce: Life Technologies Czech Republic s.r.o.
Bsp143I (Sau3AI)
8 094,90 Kč
6 690,00 Kč bez DPH
do týdne
1500 units

.

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

FastDigest enzyme available Unique buffer for 100% activity Optimal incubation at 37°C Cleavage blocked or impaired by overlapping CpG methylation Thermal inactivation at 65°C in 20 min LO certified
The Bsp143I (Sau3AI) restriction enzyme recognizes ^GATC sites and cuts best at 37°C in its own unique buffer (Isoschizomers: BfuCI, BssMI, BstKTI, BstMBI, DpnII, Kzo9I, NdeII, Sau3AIm)
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

DpnI, Bsp143I, and MboI all recognize the same sequence, but have different methylation sensitivities and cleavage sites.

For methylation sensitivity refer to product specifications.

Compatible Ends BamHI, BclI, BglII, PsuI.
Conditions for 100% Activity
  • 1X Buffer Bsp143I: 33 mM Tris-acetate (pH 7.9 at 37°C), 10 mM Mg-acetate, 66 mM K-acetate, 0.02% Triton X-100 and 0.1 mg/mL BSA
  • Incubate at 37°C
Double Digestion Perform double digestion using DoubleDigest.
Hazardous No
Isoschizomers Search for commercial isoschizomers using REsearch.
Storage Buffer Bsp143I 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
Bsp143I Buffer [Unique] 37°C 20-50 20-50 0-20 0-20 50-100 20-50 1X or 2X

Methylation Effects

  • Dam: completely overlaps - no effect.
  • Dcm: never overlaps - no effect.
  • CpG: may overlap - blocked.
  • EcoKI: never overlaps - no effect.
  • EcoBI: may overlap - no effect.
Methylation type Sequence Cleavage effect
Dam (GATC) Gm6ATC No effect
CpG 5'...GATm5C G...3'
3'...CTA Gm5C...5'		 Blocked
EcoBI (TGA(N)8TGCT) 5'...TGm6ATC(N)6 TGCT...3'
3'...AC TAG(N)6m6ACGA...5'		 No effect

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
116 0 7 22 15 15
pTZ19R/U pTZ57R pBluescriptIIKS(±) pBluescriptIISK(±) pACYC177 pACYC184
15 15 15 15 22 15

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
^GATC
  • BamHI/FastDigest BamHI (G^GATCC)
  • BclI/FastDigest BclI (T^GATCA)
  • BglII/FastDigest BglII (A^GATCT)
  • 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
^GATC GATCGATC
  • [Bsp143I (Sau3AI)/FastDigest Sau3AI (Bsp143I)]
  • Bsu15I (ClaI)/FastDigest ClaI (Bsu15I)
  • [DpnI/FastDigest DpnI]
  • [MboI/FastDigest MboI]
  • TaqI/FastDigest TaqI

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