You are fresh recruits to a molecular biology laboratory, and your new boss has tasked you to study mutations in NRAS, a gene that she suspects is involved in cancer pathogenesis. Task A: Polymerase Chain Reaction Master Mix Your first task is to isolate and amplify the NRAS gene from cDNA extracted from different samples through PCR. You will need to run a total of 7 PCRs: 3 normal samples, 3 cancerous samples, and 1 negative control. To make things easier in the lab, when running multiple reactions, the components are prepared not individually, but as a master mix—all the components for multiple reactions are prepared in bulk, except for the template DNA, which is added separately once the master mix has been distributed into individual tubes. The table below lists the different components for PCR, the available stock concentrations of these components, and the needed working concentrations for the PCR itself. Complete the table by supplying the needed volumes of each component for a single reaction and for the master mix (the first row has been filled in as an example).
You are fresh recruits to a molecular biology laboratory, and your new boss has tasked you to study mutations in NRAS, a gene that she suspects is involved in cancer pathogenesis. Task A: Polymerase Chain Reaction Master Mix Your first task is to isolate and amplify the NRAS gene from cDNA extracted from different samples through PCR. You will need to run a total of 7 PCRs: 3 normal samples, 3 cancerous samples, and 1 negative control. To make things easier in the lab, when running multiple reactions, the components are prepared not individually, but as a master mix—all the components for multiple reactions are prepared in bulk, except for the template DNA, which is added separately once the master mix has been distributed into individual tubes. The table below lists the different components for PCR, the available stock concentrations of these components, and the needed working concentrations for the PCR itself. Complete the table by supplying the needed volumes of each component for a single reaction and for the master mix (the first row has been filled in as an example).
Biochemistry
9th Edition
ISBN:9781319114671
Author:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Chapter1: Biochemistry: An Evolving Science
Section: Chapter Questions
Problem 1P
Related questions
Question
You are fresh recruits to a molecular biology laboratory, and your new boss has tasked you to
study mutations in NRAS, a gene that she suspects is involved in cancer pathogenesis.
Task A: Polymerase Chain Reaction Master Mix
Your first task is to isolate and amplify the NRAS gene from cDNA extracted from different
samples through PCR. You will need to run a total of 7 PCRs: 3 normal samples, 3 cancerous
samples, and 1 negative control. To make things easier in the lab, when running multiple
reactions, the components are prepared not individually, but as a master mix—all the
components for multiple reactions are prepared in bulk, except for the template DNA, which is
added separately once the master mix has been distributed into individual tubes.
The table below lists the different components for PCR, the available stock concentrations of these
components, and the needed working concentrations for the PCR itself. Complete the table by
supplying the needed volumes of each component for a single reaction and for the master mix
(the first row has been filled in as an example).
Transcribed Image Text:#2 Mol Bio Mutation QUESTIONS X +
← → C ✰ drive.google.com/file/d/1mcDi8k1 KtP3uaF-SQ-QgeLgnjxj20dqA/view
M Gmail ▸ YouTube
Maps
PDF #2 Mol Bio Mutation QUESTIONS.pdf
I
Escribe aquí para buscar
A good night's slee... G games Piktochart > NCBI
PCR Component
Buffer for Taq
polymerase
Forward primer
Reverse primer
dNTPs
Taq polymerase
Sterile distilled
deionized water
Template
O
Stock
concentration
8:
10X
40 uM
10 UM
25 mM
200 U/ul
n/a
Total
400 ng/ul
Total
79
Abrir con
Working
concentration
Lucid Coggle APA Diagrama
Página 1/
1X
1.0 uM
1.0 uM
1 mM
1 U/ul
(to fill reaction
volume to 10 ul)
40 ng/ul
3
Volume for 1
reaction (UL)
1.0
Q +
10.0
Simuladores y labor...
9.0 ul
63.0 ul
1.0 (added separately to each reaction
after master mix is distributed to
different tubes)
P
Volume for 7
reactions (µL)
7.0
70.0
>
Étudiant | Campus...
ESP
0 X
Actualizar
04:19 p.m.
09/08/2022
Expert Solution
This question has been solved!
Explore an expertly crafted, step-by-step solution for a thorough understanding of key concepts.
Step by step
Solved in 3 steps
Recommended textbooks for you
Biochemistry
Biochemistry
ISBN:
9781319114671
Author:
Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:
W. H. Freeman
Lehninger Principles of Biochemistry
Biochemistry
ISBN:
9781464126116
Author:
David L. Nelson, Michael M. Cox
Publisher:
W. H. Freeman
Fundamentals of Biochemistry: Life at the Molecul…
Biochemistry
ISBN:
9781118918401
Author:
Donald Voet, Judith G. Voet, Charlotte W. Pratt
Publisher:
WILEY
Biochemistry
Biochemistry
ISBN:
9781319114671
Author:
Lubert Stryer, Jeremy M. Berg, John L. Tymoczko, Gregory J. Gatto Jr.
Publisher:
W. H. Freeman
Lehninger Principles of Biochemistry
Biochemistry
ISBN:
9781464126116
Author:
David L. Nelson, Michael M. Cox
Publisher:
W. H. Freeman
Fundamentals of Biochemistry: Life at the Molecul…
Biochemistry
ISBN:
9781118918401
Author:
Donald Voet, Judith G. Voet, Charlotte W. Pratt
Publisher:
WILEY
Biochemistry
Biochemistry
ISBN:
9781305961135
Author:
Mary K. Campbell, Shawn O. Farrell, Owen M. McDougal
Publisher:
Cengage Learning
Biochemistry
Biochemistry
ISBN:
9781305577206
Author:
Reginald H. Garrett, Charles M. Grisham
Publisher:
Cengage Learning
Fundamentals of General, Organic, and Biological …
Biochemistry
ISBN:
9780134015187
Author:
John E. McMurry, David S. Ballantine, Carl A. Hoeger, Virginia E. Peterson
Publisher:
PEARSON