Draw your own PCR diagram to show the role of each component and relevance of each temperature shift during the PCR reaction.What’s happening at each of those temperatures and why is it important it changes? Important points to follow:• Samples S1 and 52 are at a suitable dilution already• PCR reactions are 'run' in the 0.2ml tubes - these should not be squeezed asthey crack.• Each PCR reactions each need to contain:ComponentGoTaq Hot Start Master Mix, 2XPrimer set (E or H; 15ng/μl)DNA to test (P1, S1 or $2; 20ng/μl)Nuclease-free waterYou will set up four reactions.95°C 5 minutes95°C 30 sec50°C 1 minLabel the tubes with details of the DNA or tube number and your workstationnumber so you know which is which next week. Once you've added the necessaryreagents, place your tubes in the correct rack at the front of the lab.We will then cycle these through the following temperatures:1xFinal Volume25.0μl40x5.0μl1.0μlup to 50μl72°C 1 min72°C 10 minThey will then be stored in the fridge (4°C) until you need them next week.

The polymerase chain reaction or PCR is a method widely used to rapidly make multiple copies of a…

Draw your own PCR diagram to show the role of each component and relevance of each temperature shift during the PCR reaction. What’s happening at each of those temperatures and why is it important it changes?

Draw your own PCR diagram to show the role of each component and relevance of each temperature shift during the PCR reaction. What’s happening at each of those temperatures and why is it important it changes?

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

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Question

Draw your own PCR diagram to show the role of each component and relevance of each temperature shift during the PCR reaction.

What’s happening at each of those temperatures and why is it important it changes?

Important points to follow:
• Samples S1 and 52 are at a suitable dilution already
• PCR reactions are 'run' in the 0.2ml tubes - these should not be squeezed as
they crack.
• Each PCR reactions each need to contain:
Component
GoTaq Hot Start Master Mix, 2X
Primer set (E or H; 15ng/μl)
DNA to test (P1, S1 or $2; 20ng/μl)
Nuclease-free water
You will set up four reactions.
95°C 5 minutes
95°C 30 sec
50°C 1 min
Label the tubes with details of the DNA or tube number and your workstation
number so you know which is which next week. Once you've added the necessary
reagents, place your tubes in the correct rack at the front of the lab.
We will then cycle these through the following temperatures:
1x
Final Volume
25.0μl
40x
5.0μl
1.0μl
up to 50μl
72°C 1 min
72°C 10 min
They will then be stored in the fridge (4°C) until you need them next week.

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