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TaqI

Kód produktu: ER0671 Kód výrobce: ER0671 Kód dodavatele: {5EDCA2B1-D0A0-42DC-A2C0-7609B3AC9F02} Výrobce: Life Technologies Czech Republic s.r.o.
2 274,80 Kč
1 880,00 Kč bez DPH
do týdne
3000 units

5'...TC G A...3'
3'...A G CT...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

Assayed using Lambda DNA (dam-) (#SD0021). TaqI is blocked by overlapping dam methylation. To avoid dam methylation, use a dam-, dcm- strain, such as GM2163. Incubation at 37°C results in 10% activity. We recommend using TaqI, HC at 37°C.

For methylation sensitivity refer to product specifications.

Compatible Ends Bsp119I, Bsu15I, Hin1I, Hin6I, HpaII, MaeII, MspI, NarI, Psp1406I, SsiI, XmiI.
Conditions for 100% Activity
  • 1X Buffer TaqI: 10 mM Tris-HCl (pH 8.0 at 37°C), 5 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 TaqI is supplied in: 10 mM Tris-HCl (pH 7.5 at 25°C), 300 mM KCl, 1 mM DTT, 0.1 mM EDTA, 0.5 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
TaqI Buffer (Unique) 65°C 0-20 20-50 20-50 20-50 20-50 20-50 1X or 2X

Methylation Effects

  • Dam: may overlap - blocked.
  • Dcm: never overlaps - no effect.
  • CpG: completely overlaps - no effect.
  • EcoKI: never overlaps - no effect.
  • EcoBI: never overlaps - no effect.
Methylation type Sequence Cleavage effect
Dam(GATC) 5'...TCGm6A TC...3'
3'...AGC Tm6AG...5'
Blocked
CpG TCGA No effect

Cleavage efficiency close to the termini of PCR fragments

bp from the recognition site to fragment end
1 2 3 4 5
0 20-50 50-100

Number of recognition sites in DNA molecules

Lambda ΦX174 M13mp18/19 pBR322 puc18/19 pUC57
121 10 12 7 4 4
pTZ19R/U pTZ57R pBluescriptIIKS(-/+) pBluescriptIISK(-/+) pACYC177 pACYC184
5 5 7 7 10 10

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^CGA
  • BmeT110I* (CY^CGAG)
  • Bsp119I (BstBI)/FastDigest Bsp119I (TT^CGAA)
  • Bsu15I (ClaI)/FastDigest ClaI (Bsu15I) (AT^CGAT)
  • XmiI (AccI)/FastDigest AccI (XmiI)* (GT^CGAC)
  • TaqI/FastDigest TaqI

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^CGA TCGCGA
  • Bsh1236I (BstUI)/FastDigest Bsh1236I
  • Hpy188III
  • Bsp68I (NruI)/FastDigest NruI (RruI)

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