Question: you want to set up pcr reactions with varying amounts...
You want to set up PCR reactions with varying amounts of magnesium to determine the effects of magnesium ion concentration on the yield and quality of PCR product. You plan to use the 2X GoTaq Mastermix (see manual for components), 1ng of your genomic DNA (at 30ng/ul) and 2 ul of the forward/reverse primer mix at 5 uM each. You have a 75mM stock of magnesium chloride and you want a working concentration of 4mM in your PCR reaction. Your final PCR volume is 40 ul. What volume of each of the following stock solutions will you mix to make your reaction?
Could someone please guide me through the steps of how to solve this PCR problem? I'm at a loss as to where to start.
EDIT: From the manual: "For regular PCR we use a reagent called GoTaq Green Master Mix 2X to do the PCR: the Go Tag contains the following:
Buffer (pH 8.5)
400 uM dATP, 400uM dGTP, 400 uM dCTP, 400 uM dTTP
Taq DNA Polymerase
EDIT 2: Use the working concentration of DNA = 1ng of 30ng/ul and use the stock concentration 75mM for MgCl2