What is Herculase?
What is Herculase?
Herculase® is a DNA Polymerase designed to deal with difficult to amplify targets such as long or GC-rich DNA templates. This combination amplifies a broad range of DNA lengths (0.1 – 48 kb) with reportedly greater yield and higher fidelity than Taq DNA polymerase alone.
What are the buffers used in PCR?
PCR is carried out in a buffer that provides a suitable chemical environment for activity of DNA polymerase. The buffer pH is usually between 8.0 and 9.5 and is often stabilized by Tris-HCl. For Taq DNA polymerase, a common component in the buffer is potassium ion (K+) from KCl, which promotes primer annealing.
How many buffers are in PCR?
PCR Page 4 Polymerase Chain Reaction, 12/2004 4 buffers are often available in 10X concentration and are sometimes Taq formulation-specific. Although most protocols recommend a final buffer concentration of 1X, increasing the concentration to 1.5X might result in increased PCR product yield.
Why buffer is used in PCR?
Why KCl is used in PCR?
The KCl salt in the PCR buffer acts by neutralizing the charge present on the backbone of DNA. During the elongation step of the PCR, the primer has to anneal or stick properly to the template and this is facilitated by the KCl.
Which chemical is used in PCR?
The key ingredients of a PCR reaction are Taq polymerase, primers, template DNA, and nucleotides (DNA building blocks). The ingredients are assembled in a tube, along with cofactors needed by the enzyme, and are put through repeated cycles of heating and cooling that allow DNA to be synthesized.
What is PCR and its application?
Polymerase chain reaction (PCR) is a technique used to exponentially amplify a specific target DNA sequence, allowing for the isolation, sequencing, or cloning of a single sequence among many. PCR was developed in 1983 by Kary Mullis, who received a Nobel Prize in chemistry in 1993 for his invention.
What 5 components are needed for PCR?
Why is the PCR reaction buffer viscous in nature?
It is viscous in nature which prevents the binding or sticking of Taq DNA polymerase and other PCR reaction reagents to the wall of the PCR tube. By doing this, it provides more surface area to the reaction between different chemicals. Also, it prevents excess evaporation during extreme heating in the PCR.
How is formamide used in the PCR reaction buffer?
Formamide increases the PCR amplification efficiency of the GC rich region too. A concentration of 1% to 10% is preferable. Notably, as the formamide can easily form primer-dimer, use it if needed and based on the GC content of the template. Once the DNA template denatured, no hydrogen bonds are present between bases.
Which is the best detergent for the PCR reaction buffer?
However, apart from SDS and CTAB, other non-ionic detergents like Triton X100, Nonidet P40, and Tween20 are great PCR enhancers. Using these detergents in the PCR reaction buffer can greatly increase the yield and efficiency of the PCR reaction.
Why is EDTA used in the PCR reaction buffer?
Using EDTA into PCR will chelate the excess unused Mg2+ ions that might inhibit the PCR, also, it chelates other ions presents into the PCR reaction mixture. Therefore, if we use EDTA in a lesser proportion of MgCl2, it will enhance the PCR reaction.