How can tween 20 affect PCR?

As little as 0.01\% SDS contamination of the template DNA (left-over from the extraction procedure) can inhibit PCR by reducing Taq polymerase activity to as low as 10\%, however, inclusion of 0.5\% Tween-20 or -40 will effectively neutralize this effect.

What is the role of DMSO in PCR?

DMSO binds to the DNA and prevents the reannealing of single-stranded DNA. It also facilitates the annealing of primer with a templet. Therefore, it increases the specificity and yield of the PCR reaction. Generally, the GC content of the template DNA for PCR is between 45\% to 52\%.

Does tween interfere with PCR?

Ionic detergents (e.g. sodium deoxycholate, sarkosyl and SDS) are highly inhibitory for the PCR, whereas non-ionic detergents (e.g. Nonidet P-40, Tween 20, Triton X-100 and N-octyl glucoside) cause PCR inhibition only at relatively high concentrations (Weyant et al. 1990).

Why is BSA used in PCR?

Thermo Scientific Bovine Serum Albumin (BSA) is ideal for stabilization of enzymes during storage and for enzymatic reactions where the absence of nucleases is essential. BSA increases PCR yields from low purity templates. It also prevents adhesion of enzymes to the reaction tubes and tip surfaces.

What is the role of KCl in PCR?

The KCl salt in the PCR buffer acts by neutralizing the charge present on the backbone of DNA. Thus, it functions by reducing the repulsion between the negatively charged DNA strands i.e. the primer and template, thereby stabilizing the primer-template binding.

Why buffer is used in PCR?

PCR buffer is necessary to create optimal conditions for activity of Taq DNA polymerase. Buffers often contain Tris-Hcl, KCl, and sometimes MgCl2. PCR Page 4 Polymerase Chain Reaction, 12/2004 4 buffers are often available in 10X concentration and are sometimes Taq formulation-specific.

What does GC enhancer do?

The 360 GC Enhancer is used for difficult-to-amplify templates, especially for templates with high GC content or GC-repeats. The enhancer can also increase specificity in reactions that generate non-specific products for those templates which have a localized GC-rich sequence.

Can BSA inhibit PCR?

BSA is widely used as a PCR inhibition alleviating agent in microfluidic devices in both static11–15 and dynamic passivation modes. Also, there are many reports, where combinations of static and dynamic surface passivation using BSA have been applied to PCR micro-devices.

How do you get 5m betaine?

Q-solution (5 M betaine) Dissolve 29.275 g betaine in final volume of 50 ml sterile ddH2O. Sterilize by 0.22 um filter, and store at -20¡ãC.