Answered: Make the PCR Cocktail I field out my…

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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Make the PCR Cocktail

I field out my work.. but I don't know which one is incoeert..

This table lists the ingredients, stock reagent concentrations, and concentrations in the PCR reaction.

Prepare a "PCR cocktail" to be added to your samples to achieve these concentrations. Make enough cocktail to run nine samples.

[Four student samples + three positive controls + one negative control + one extra.]

{Hint: Remember that the concentration in the reaction is not the same as the concentration in the cocktail!]

Component	Stock Concentration	Concentration in the PCR reaction	Volume per reaction	Volume to make cocktail
Sterile water	-	-	0µl	0µl
PCR buffer w/ MgCl₂	10x	1x	4µl	36µl
Nucleotide mix	10 mM	0.2 mM	0.8µl	7.2µl
Primer 1 (Forward)	10 µM	1.0 µM	4µl	36µl
Primer 2 (Reverse)	10 µM	1.0 µM	4µl	36µl
Taq DNA polymerase	5 U/µl	1.0 U	8µl	72µl
DNA template (sample)	-	~1 ng	20 µl	180µl
Total	-	-	40 µl	360 µl

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