1. What is a “dNTP” and why are they included in the reaction mixture? 2. What will be the final concentration of 25 mM MgCl2, 5 mM dNTPs, and each 25 µM primer in one of the reaction tubes with the final volume of 50 microliters? Hint: use the information in the left column of Table 1 to help you with these calculations. (For one reaction) item  5.25x master mix – add things in order 32.7 µl  ddH2O  171.7 µl 10 µl  5x reaction buffer  52.5 µl 3 µl  25 mM MgCl2  15.75 µl  0.5 µl  5 mM dNTPs  2.63 µl  1 µl  25 µM “forward” primer (#3012) 5.25 µl  1 µl 25 µM “reverse” primer (#3013)  5.25 µl 0.8 µl thermostable polymerase 4.2 µl (do not add until needed)       Total: 49 µl   Total: 257.3 µl

Human Anatomy & Physiology (11th Edition)
11th Edition
ISBN:9780134580999
Author:Elaine N. Marieb, Katja N. Hoehn
Publisher:Elaine N. Marieb, Katja N. Hoehn
Chapter1: The Human Body: An Orientation
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Problem 1RQ: The correct sequence of levels forming the structural hierarchy is A. (a) organ, organ system,...
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1. What is a “dNTP” and why are they included in the reaction mixture?

2. What will be the final concentration of 25 mM MgCl2, 5 mM dNTPs, and each 25 µM primer in one of the reaction tubes with the final volume of 50 microliters? Hint: use the information in the left column of Table 1 to help you with these calculations.

(For one reaction) item  5.25x master mix – add things in order
32.7 µl  ddH2O  171.7 µl
10 µl  5x reaction buffer  52.5 µl
3 µl  25 mM MgCl2  15.75 µl
 0.5 µl  5 mM dNTPs  2.63 µl
 1 µl  25 µM “forward” primer (#3012) 5.25 µl
 1 µl 25 µM “reverse” primer (#3013)  5.25 µl
0.8 µl thermostable polymerase 4.2 µl (do not add until needed)     
 Total: 49 µl   Total: 257.3 µl
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