|
At Bioline we pride ourselves in supplying high-quality polymerases to suit your requirements. To aid your selection of the most suited enzyme for your specific applications, please see our polymerase selection guide.
All our polymerases are guaranteed for a period of 12 months from the date of purchase. These should be stored at
-20°C during this time for optimal retention of activity.
Please Note: We do not recommend the storage of our polymerases at -80°C as ice crystals could form on the active site, which may affect or destroy the activity of the enzyme.
PCR can be a challenging technique, with various parameters to optimize to achieve the best results. If you are having problems, these could be easily resolved by addressing a few issues. Please see our PCR troubleshooting guide for suggestions and help with your specific problems.
These terms refer to parameters to be considered when performing PCR, and are important features in selecting the correct enzyme for your needs. Understanding what they mean is therefore crucial:
- Yield: The amount of DNA produced in a PCR reaction
- Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand.
- Processivity: The length of time a polymerase is associated with the template, and therefore the size of fragment which can be amplified.
- Specificity: A measure of the unwanted by-products generated in a reaction.
In nature, the ability of a DNA polymerase to correct misincorporations of nucleotides in the DNA strand being elongated is often crucial to the survival of the host organism. This ability is termed “proofreading activity”, and occurs in the 3’ to 5’ direction. This activity also leads to the polymerase removing unpaired nucleotides overhanging at the 3’end (A-overhangs), creating blunt ends.
Please see the following table for the appropriate error rates:
|
|
BIOLASE, IMMOLASE, Mango Taq, MyTaq DNA Polymerase
, Taq DNA Polymerase.
|
1.1 x 104 base substitutions/bp (Tindall and Kunkel, 1988) |
| 2.4 x 105 frameshift mutations/bp (Tindall and Kunkel, 1988) |
| 2.1 x 104 errors/bp (Keohavang and Thilly, 1989) |
| 7.2 x 105 errors/bp (Ling et al., 1991) |
| 8.9 x 105 errors/bp (Cariello et al., 1991) |
| 2.0 x 105 errors/bp (Lundberg et al., 1991) |
| 1.1 x 104 errors/bp (Barnes, 1992) |
| ACCUZYME |
1.6 x 106 errors/base (Lundberg et al., 1991). |
| VELOCITY |
4.4 x 10-7 errors/base (Frey & Suppmann, 1995). |
One unit is defined as the amount of enzyme that incorporates 10nmoles of dNTPs into acid- insoluble form in 30 minutes at 72°C.
Yes, BIO-X-ACT Long and Short are highly suited to amplification from difficult or problematic templates, such as high GC% regions.
BIO-X-ACT Short will process problematic templates up to 5Kb in length.
BIO-X-ACT Long is specifically formulated for the amplification of long fragments, and will extend Genomic DNA of up to 20Kb, and Lambda DNA of up to 30Kb.
The fidelity of these two polymerases is 17-fold higher than that of Taq, and they will extend at 30-45s/Kb.
The BIO-X-ACT range of polymerases require a lot of MgCl2 in the reaction, at times as much as twice more than conventional enzymes. This is an important parameter for successful PCR, and a titration of MgCl2 is recommended for ascertaining optimal PCR results. A vial of 50mM MgCl2 Solution is provided with all our polymerases for your convenience. |
