How will you modify the reaction if the given 10x reaction buffer contains no MgCl2 and a tube of 25mM MgCl2 is provided separately?
What is the role of reaction buffer??
2006-10-06 18:39:18 · 3 answers · asked by snowynite19 1 in Science & Mathematics ➔ Biology
"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
2006-10-07 01:45:04 · answer #1 · answered by bellerophon 6 · 0⤊ 0⤋
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??
2006-10-07 04:10:06 · answer #2 · answered by Davo 2 · 0⤊ 0⤋
ahhhh, just reading your question reminds me of the days when I took classes when I had to know that stuff! :) I'm sorry I can't remember that info right off, it would take some delving into my old class notes - I hope someone remembers!
2006-10-07 02:47:07 · answer #3 · answered by natureutt78 4 · 0⤊ 0⤋