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
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
Section: Chapter Questions
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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