PCR Master
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| for routine amplification of up to 3 kb genomic DNA targets Cat. No.
1 636 103 (100 reactions) |
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| Description |
The PCR Master Kit contains 10 vials of 2x concentrated PCR reaction
mix (Taq DNA Polymerase, reaction buffer with MgCl2, stabilizing
detergent, dNTPs) plus water PCR grade (for dilution), sufficient for 100
PCR reactions. Taq DNA Polymerase was originally isolated from the thermophilic
eubacterium Thermus aquaticus BM. It is now supplied as a recombinant enzyme
from E. coli. The enzyme is highly purified and is free of nonspecific endo-
or exonucleases. Taq DNA Polymerase consists of a single polypeptide chain
with a molecular weight of approximately 95 kD. It is a highly processive
5'–3' DNA polymerase which lacks 3'–5' exonuclease activity.
Note: For additional convenience and reliable PCR with minimum reagent preparation,
we offer the PCR
Master [Cat. No. 1636103, 2x conc. PCR reaction mix (Taq DNA Polymerase,
reaction buffer with MgCl2, stabilising detergent, dNTPs)] plus water PCR
grade for dilution. Further the PCR
Core Kit (Cat. No. 1578553), containing Taq DNA Polymerase, dNTP
stock solution, PCR reaction buffer with and without MgCl2 and
a MgCl2 stock solution. |
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| Application |
Amplifies genomic targets up to 3 kb with good yield and specificity.
Capable of amplifying lambda DNA up to 10 kb. The enzyme is ideal for:
- Routine amplification of single-copy genes from eukaryotic genomes
- Labeling of PCR products with modified nucleotides (DIG-dUTP, biotin-dUTP,
fluorescein-dUTP)
- Prevention of carry-over contamination, by incorporation of dUTP
and degradation of contaminating DNA with Uracil-DNA Glycosylase
- Cycle sequencing
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| Operating parameters |
- pH optimum: approx. 9 (adjusted at 20°C)
- Temperature optimum (for elongation): approx. 75°C
Note: The DNA polymerase has a half life at 95°C of approx. 40 min
- Divalent ion requirement: Mg2+ (standard concentration,
1.5 mM)
- dNTP requirement: approx. 200 µM for each dNTP
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| Effects of additives |
| Tween 20 (0.5–1.0%) detergent has been used to enhance the efficiency
of Taq DNA Polymerase in certain PCRs. Other additives reported to have
enhancing effects include DMSO, gelatine, glycerol, betaine, spermidine,
T4 gene 32 protein, BSA, and ammonium sulfate. |
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| Key advantages |
- High PCR yield, because it is stable during prolonged, repetitive
high temperature incubations
- Enhances specificity of amplification, because it is highly processive
and free of exonuclease and nicking activities
- Increases sensitivity, because it produces good yields even when
template amounts are limiting
- Best results, because it has proved superior to competitors’ Taq
DNA Polymerase preparations (see "Not
all Taq DNA polymerase preparations are the same")
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