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7.You want to set up a PCR reaction. The protocol suggests a 50 µL reaction volume...

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Biology

7.You want to set up a PCR reaction.The protocol suggests a 50 µL reaction volume containing 1X buffer,200 µM dNTPs, 0.5 µM each primer (forward and reverse), 250 ngtemplate DNA, and 1 U enzyme.

You have the following reagents: 5 Xbuffer, 10 mM dNTPs, 10 µM each primer (forward and reverse), 125ng/µL template DNA, and 2 U/µL enzyme.

How many µL of water, reactionbuffer, dNTPs, each primer, template DNA and enzyme would youcombine to set up that reaction? (note that the order I’ve listedthese components is the order you’d add them to the reaction)

Answer & Explanation Solved by verified expert
4.1 Ratings (568 Votes)
Ans I Buffer Using C1V1 5X stock buffer C2V2 final PCR mix 1X 50 uL Or 5X x V1 1X x 500 uL Or V1 1X x 500 uL 5X Hence V1 100 uL Hence required volume of 5X stock buffer 100 uL II dNTPs Using C1V1 10 mM    See Answer
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