Taq DNA Polymerase
|
for routine amplification of up to 3 kb genomic
DNA targets Cat. No. 1 647 679 250 units (1 unit/µl)
Cat. No. 1 647 687 4 x 250 units (1 unit/µl)
Cat. No. 1 146 165 100 units (5 units/µl)
Cat. No. 1 146 173 500 units (5 units/µl)
Cat. No. 1 418 432 4 x 250 units (5 units/µl)
Cat. No. 1 596 594 10 x 250 units (5 units/µl)
Cat. No. 1 435 094 20 x 250 units (5 units/µl) |
| |
| Description |
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. |
| |
| 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
(see also "Not
all Taq DNA polymerase preparations are the same") |
| |
Application profile
|
 |
| |
| 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
|
| |
| Effect 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. |
| |
| Experimental result |
| |
 |
| |
| Five different lots of Taq DNA Polymerase have
been tested in PCR to amplify a 0.5 kb fragment of lambda DNA. Reliable,
consistent results were obtained with every lot of Roche Applied Science
Taq DNA Polymerase that was tested. |
| |
| |
| 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")
|