LaboPass™ IP-Taq DNA Polymerase is a thermostable DNA polymerase cloned from Thermus aquaticus and a recombinant form expressed in E.coli. This enzyme possesses 5' to 3' exonuclease activity, but lacks a 3' to 5' exonuclease proofreading activity. The enzyme purified with high purity contains a very low level of contaminating E.coli DNA, which minimizes false-positive results, especially when the amplicon is bacterial sequence (e.g. 16S rRNA).
- Applications
- - General PCR for detection
- - Colony PCR
- - Real-time PCR
- - A-tailing for TA-cloning
Supplied reagents
LaboPass™ IP-Taq DNA Polymerase is provided with an optimized buffer to improved PCR yield.
- 10X IP-Taq buffer I and II (with MgCl2)
- LaboPass™ IP-Taq DNA Polymerase is supplied with two types of reaction buffer with different salt formulation. Generally, Buffer I works well in most PCR reaction. The use of Buffer II can be tried if PCR products are not satisfactory (nonspecific, little or no products) using Buffer I.
- 5X Tuning buffer
- Tuning buffer can improve PCR efficiency in reaction using problematic template DNA containing high GC contents or stable secondary structure. Thus, it is advantageous to amplify complicated long target sequences.
| Component | Volume |
|---|---|
| IP-Taq polymerase (2.5 unit/μl) | 250 unit X 2 |
| 10X IP-Taq buffer I (with MgCl2) | 1 ml X 2 |
| 10X IP-Taq buffer II (with MgCl2) | 1 ml |
| dNTPs (each 2.5 mM) | 500 μl X 2 |
| 5X Tuning buffer | 1 ml |
High amplification efficiency
The PCR amplification efficiency was compared with other commercial Taq polymerase. LaboPass™ IP-Taq polymerase shows comparable or superior quality.
Lane M : 1 kb Labo DNA ladder
Lane M : 1 kb Labo DNA ladder
PCR performance
Various sizes of PCR produces can be amplified using LaboPass™ IP-Taq DNA Polymerase.
Lane M : 1kb Labo DNA ladderDNA template : λ DNA
| Standard Reaction (50 µl) | Volume |
|---|---|
| 10X IP-Taq buffer I or II | 5 μl |
| dNTPs (each 2.5 mM) | 4 μl |
| 5X Tuning Buffer | 10 μl (optional) |
| Forward Primer | 10~50 pmoles |
| Reverse Primer | 10~50 pmoles |
| DNA Template | Variable * |
| IP-Taq Polymerase | 1 μl |
| Distilled Water | up to 50 μl |
Quality control
Each lot of IP-Taq polymerase, 10X IP-Taq buffer and dNTPs is tested for contamination such as E.coli genomic DNA, nicking, endo-nuclease and exo-nuclease.
- Test for nuclease activity
- Nicking, endonuclease and exonuclease activity were not detected after the incubation of 0.5 μg of supercoiled pUC19, λ DNA or HindIII digested λ DNA with 10 units of this enzyme for 4 hour at 37°C or 72°C.
- Test for E.coli genomic DNA contamination
- When compared with a competitor's DNA polymerase, LaboPass™ IP-Taq polymerase verified to have noE.coli genomic DNA contamination.
Lane M : 1 kb Labo DNA ladderLane PC : Positive control (E.coli gDNA, 10 ng)