10X Reaction Buffer for phi29 DNA Polymerase

Kód produktu: B62 Kód výrobce: B62 Kód dodavatele: {473C60C0-8A97-46F4-8244-8141CEB0E6B2} Výrobce: UAB Fermentas
548,13 Kč
453,00 Kč bez DPH
do týdne
5x1 ml
Recombinant enzyme Thermal inactivation at 65°C in 10 min
phi29 DNA Polymerase is a highly processive polymerase with strong strand displacement activity that allows for highly efficient isothermal DNA amplification.
Zobraz detailní popis

Detailní popis

Thermo Scientific phi29 DNA Polymerase is a highly processive polymerase (up to more than 70 kb) featuring strong strand displacement activity, which allows for highly efficient isothermal DNA amplification (see Reference 1). phi29 DNA Polymerase also possesses a 3'→5' exonuclease (proofreading) activity acting preferentially on single-stranded DNA (see Reference 2) or RNA (see Reference 3). Therefore 3'-modified primers are highly recommended (see Reference 4).

Highlights

  • Highest processivity and strand displacement activity among known DNA polymerases – more than 70 kb long DNA stretches can be synthesized (see Reference 1)
  • Highly accurate DNA synthesis (see Reference 5)
  • Extremely high yields of amplified DNA even from minute amounts of template
  • Amplification products can be directly used in downstream applications (PCR, restriction digestion, SNP genotyping, etc.)

Applications

  • Rolling circle amplification (RCA) (see Reference 6): generation of periodic DNA nanotemplates (see Reference 7)
  • Multiple displacement amplification (MDA) (see Reference 8)
  • Unbiased amplification of whole genome (WGA, see Figure 1 in Supporting Data):
    • amplification of DNA for SNP (see Reference 9) and STR (see Reference 10) detection
    • cell-free amplification of DNA from single cells (see References 11, 12)
    • pathogenic organisms or metagenomes (see Reference 13)
    • amplification of DNA from filter paper blood spot samples (see Reference 14)
  • DNA template preparation for sequencing
  • Protein-primed DNA amplification (see Reference 15)
  • In situ genotyping with padlock probes (see Reference 17)
  • Recombination based-cloning (see Reference 18)
  • Cell-free cloning of lethal DNA (see Reference 19)
  • RNA-primed DNA amplification (see Reference 16)

Note

Addition of Pyrophosphatase to the reaction mixture with phi29 DNA Polymerase may enhance DNA synthesis (see Reference 8).

Use of this enzyme in certain applications may be covered by patents and may require a license.

10× Reaction Buffer 330 mM Tris-acetate (pH 7.9 at 37°C), 100 mM Mg-acetate, 660 mM K-acetate, 1% (v/v) Tween 20, 10 mM DTT.
Definition of Activity Unit
  • One unit of the enzyme catalyzes the incorporation of 0.5 pmol of dCMP into a polynucleotide fraction (adsorbed on DE-81) in 10 min at 30°C.
  • Enzyme activity is assayed in the following mixture: 50 mM Tris-HCl (pH 7.5), 10 mM MgCl2, 1 mM DTT, 0.01 mg/mL lambda DNA/HindIII, 0.2 µM dCTP, including [3H]-dCTP, 0.2 mM dATP, 0.2 mM dGTP, and 0.2 mM dTTP.
Hazardous No
Inactivation Inactivated by heating at 65°C for 10 min.
Inhibition Inhibitors: aphidicolin, N2-(p-n-butylphenyl)-dGTP (BuPdGTP), 2-(p-n-butylanilino)-dATP (BuAdATP) (20).
Molecular Weight 66.7 kDa monomer
Quality Control The absence of endodeoxyribonucleases confirmed by appropriate quality tests.
Source E.coli cells with a cloned gene 2 of Bacillus subtilis phage phi29.
Specific Activity 100,000 U/mg.
Storage Buffer The enzyme is supplied in 50 mM Tris-HCl (pH 7.5), 0.1 mM EDTA, 1 mM DTT, 100 mM KCl, 0.5% (v/v) Nonidet P40, 0.5% (v/v) Tween 20, and 50% (v/v) glycerol.
Storage Condition -20 C

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