what is the importance or the function of MgCl2 in PCR????

Question

any recommended reading

Answer 1

two things....

The polymerase requires a divalent cation (in this case Mg++ from the MgCl2) in order to function.

The Mg++ also helps stablize the two strands. A normal concentration of Mg++ in the reaction is 2.5uM. Higher concentrations yield greater promiscuity of the polymerase (ie false priming). Lower concentrations of Mg++ increase specificity of the target you're trying to amplify (ie. more stringent and exact matching of bp's for priming and amplification to occur).

Answer 2

Mgcl2 Pcr

Answer 3

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RE:
what is the importance or the function of MgCl2 in PCR????
any recommended reading

Answer 4

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"Why is magnesium so important in PCR?Magnesium affects DNA polymerase activity, which can affect yield; and primer annealing, which can affect specificity. The dNTPs and template bind magnesium and reduce the amount of free magnesium available for enzyme activity. Higher concentration of free magnesium can result in greater yield but can also increase non-specific amplification and reduce fidelity." (from the source below) Well you need steady pH at the optimum value of the enzyme, so that's why in all enzynmatic reactions you hav buffers. Sometimes they contain extra ingredients to maximize the enzyme's performance like salt for enzymes which require high ionic strength

Answer 5

Not 100% sure, but I believe the Magnesium Chloride helps to protect the DNA from breaking apart, and being completely denatured. When placed into a thermal cycler, where there are larger heat fluctuations, you have to remember to be careful with the DNA. I think the interaction between MgCl2 and DNA has to do with charges in the major and minor groove??

Answer 6

simply, Mg serves a a co-factor for your taq polymerase.

Enzymes sometimes require co-factors or coenzymes to function.