Complete the following tasks. 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). PCR Component Buffer for Taq polymerase Forward primer Reverse primer dNTPs Taq polymerase Sterile distilled deionized water Template Stock concentration 10X 40 uM 10 uM 25 mM 200 U/ul n/a Total 400 ng/ul Total Working concentration 1X 1.0 uM 1.0 um 1 mM 1 U/ul (to fill reaction volume to 10 ul) 40 ng/ul Volume for 1 reaction (µl.) 1.0 Volume for 7 reactions (μl) 10.0 7.0 9.0 ul 63.0 ul 1.0 (added separately to each reaction after master mix is distributed to different tubes) 70.0
Complete the following tasks. 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). PCR Component Buffer for Taq polymerase Forward primer Reverse primer dNTPs Taq polymerase Sterile distilled deionized water Template Stock concentration 10X 40 uM 10 uM 25 mM 200 U/ul n/a Total 400 ng/ul Total Working concentration 1X 1.0 uM 1.0 um 1 mM 1 U/ul (to fill reaction volume to 10 ul) 40 ng/ul Volume for 1 reaction (µl.) 1.0 Volume for 7 reactions (μl) 10.0 7.0 9.0 ul 63.0 ul 1.0 (added separately to each reaction after master mix is distributed to different tubes) 70.0
Human Heredity: Principles and Issues (MindTap Course List)
11th Edition
ISBN:9781305251052
Author:Michael Cummings
Publisher:Michael Cummings
Chapter12: Genes And Cancer
Section: Chapter Questions
Problem 13QP: The search for the BRCA1 breast cancer gene discussed in this chapter was widely publicized in the...
Related questions
Question
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).
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 with 11 images
Knowledge Booster
Learn more about
Need a deep-dive on the concept behind this application? Look no further. Learn more about this topic, biology and related others by exploring similar questions and additional content below.Recommended textbooks for you
Human Heredity: Principles and Issues (MindTap Co…
Biology
ISBN:
9781305251052
Author:
Michael Cummings
Publisher:
Cengage Learning
Human Heredity: Principles and Issues (MindTap Co…
Biology
ISBN:
9781305251052
Author:
Michael Cummings
Publisher:
Cengage Learning