Buffers
Bst DNA Polymerase is an enzyme derived from the large fragment of Bacillus stearothermophilus DNA Polymerase I. It contains 5´- 3´ DNA polymerase activity and strong strand displacement activity but lacks 5´- 3´ exonuclease activity. The strong strand displacement activity enables Bst DNA Polymerase to synthesize DNA at a constant temperature making it an ideal enzyme for isothermal amplification, including HDA, MCA, and for Loop-Mediated Isothermal DNA Amplification (LAMP).
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High Conc. Glycerol-Free Bst, MDX018
High concentration glycerol-free DNA polymerase (exo-), with strand-displacement properties. used for Isothermal DNA amplification such as LAMP (Loop-mediated Isothermal Amplification).
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Inhibitor-Tolerant Bst Buffer, MDX019
Inhibitor-Tolerant Bst Buffer is specially designed to overcome the inhibition present in samples in direct LAMP reactions.
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Bst Reaction Buffer, 10x, MDX076
Reaction Buffer optimized for use with Bst DNA Polymerase.
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Enzyme Dilution Buffer, 1x, MDX078
Glycerol containing1x dilution buffer, for the dilution of enzymes to reaction concentration.
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Enzyme Dilution Buffer, 10x (Glycerol free), MDX080
Glycerol-free,10x dilution buffer, for the dilution of enzymes to reaction concentration.
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Isothermal amplification such as loop-mediated isothermal amplification (LAMP) is a DNA amplification technique that can be performed at a single temperature. LAMP is currently considered one of the most powerful isothermal amplification techniques, relying on a strand-displacement polymerase combined with four to six primers. These primers recognize several specific regions in the target DNA and two of the primers form loop structures to facilitate subsequent rounds of amplification producing high levels of DNA.
Meridian’s Bst polymerases have strong strand displacement activity, fast polymerization, and enhanced inhibitor and salt tolerance when used with the specialized Inhibitor-Tolerant reaction buffer. The 100x high-concentration, glycerol-free BST enzyme was specifically designed to maximize assay flexibility for LAMP diagnostic test developm
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FAQs: Taq HS DNA Polymerase
If my PCR testing is being impeded by too many PCR inhibitors, what can I do for optimization?
What do the terms yield, fidelity, processivity and specificity actually mean?
These terms refer to parameters to be considered when performing PCR testing.
Yield: The amount of DNA produced in a PCR reaction. High yield increases sensitivity.
Fidelity: The accuracy of the enzyme at incorporating the correct dNTP to the elongating DNA strand. For standard PCR this is not important, but for sequencing or expression a high-fidelity mix (such as High-Specificity Pfu HS Mix (MDX006)) may be required.
Processivity: The length of time a polymerase is associated with the template and therefore the size of fragment which can be amplified (the amplicon). Taq HS DNA Polymerase will work with amplicons up to 5 kb.
Specificity: A measure of the unwanted by-products generated in a reaction. This is the reason for having hot start PCR, this prevents miss-priming at low temperatures and non-specific amplification
What applications can Taq DNA Polymerase be used for?
Taq DNA Polymerase has been validated with a broad range of PCR templates including DNA extracted from human, animal and plant samples, making it the ideal choice for the following applications:
• End-point PCR testing
• High-yield PCR testing
• Fast PCR testing
• Multiplex PCR testing
• qPCR and RT-qPCR testing
Can Taq DNA Polymerase be used for qPCR?
Yes, Taq DNA Polymerase can be used for quantitative PCR amplification (qPCR), but it will require a hot-start antibody (MDX014), qPCR buffer such as Fast qPCR Buffer, 4x (MDX033), or for RT-qPCR, a one-step buffer such as 1-Step qPCR Buffer, 4x (MDX034) and reverse transcriptase such as MMLV-RT (MDX044).
How fast is Taq DNA Polymerase?
This depends on the length of the amplicon and the complexity of the template. For fast PCR, the initial denaturation will remain 1 minute, but with low complexity template such as plasmid DNA, an annealing of 5 seconds and extension time of 10 seconds is sufficient for amplicons up to 1 kb. In order to find the fastest optimal condition, for more complex or longer amplicons, we suggest incrementally increasing the extension time successively up to 30 s/kb.
Can Taq Polymerase be used for difficult samples like GC rich or bisulfite converted DNA?
Yes, Taq DNA Polymerase works very well with difficult samples like GC rich or bisulfite converted DNA. However a hot-start to the PCR (such as MDX014) is important, as it decreases primer dimer and unspecific amplification.
If optimization is required, we would suggest starting with the annealing temperature and with bisulfite conversion, the process can fragment the DNA, so it may be useful to increase the amount of template and polymerase.
What is the error rate of Taq DNA Polymerase?
The enzyme has an error rate of approximately 1 error per 2.0 x 10⁵ nucleotides incorporated. If you are looking for a DNA polymerase with a lower error rate, for next-generation sequencing (NGS) library or expression a high-fidelity DNA polymerase (such as High-Fidelity Pfu (MDX003)) or mix (such as High-Specificity Pfu HS Mix (MDX006)) should be used.
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