100% satisfaction guarantee Immediately available after payment Both online and in PDF No strings attached
logo-home
PCR and DNA Sequencing Exam Questions and Answers Already Passed $7.99   Add to cart

Exam (elaborations)

PCR and DNA Sequencing Exam Questions and Answers Already Passed

 2 views  0 purchase
  • Course
  • PCR and DNA Sequencing
  • Institution
  • PCR And DNA Sequencing

PCR and DNA Sequencing Exam Questions and Answers Already Passed Polymerase Chain Reaction (PCR) - Answers Is very fluid and changing; always new tech. Developed in the 1980s. Reiterative DNA synthesis in vitro using specific oligonucleotide primer pairs. Short stretch of nucleotide, usua...

[Show more]

Preview 2 out of 6  pages

  • October 21, 2024
  • 6
  • 2024/2025
  • Exam (elaborations)
  • Questions & answers
  • PCR and DNA Sequencing
  • PCR and DNA Sequencing
avatar-seller
TutorJosh
PCR and DNA Sequencing Exam Questions and Answers Already Passed

Polymerase Chain Reaction (PCR) - Answers Is very fluid and changing; always new tech.



Developed in the 1980s. Reiterative DNA synthesis in vitro using specific oligonucleotide primer pairs.



Short stretch of nucleotide, usually 20 nucleotides long used as primers for DNA synthesis.



exponential amplification of a specific genomic DNA fragment. MANY MANY copies.

Tube requirements - Answers Template DNA (any type of DNA)



DNA polymerase



dNTP's (dATP, dCTP, dGTP)



Buffer, right amount of salt



A pair of single stranded Oligonucleotide Primers that hybridize to a specific genomic locus (position)
need pairs. (5' to 3') and (3' to 5').

Reaction requirements - Answers the reaction is run for many cycles, each cycle has three steps:



Denature 95C

Anneal 60C

Polymerization 72C



Typically run for 30-40 cycles

Denature - Answers separate the strands.

, anneal primers - Answers Lower temp to create H-bonds between primers and template strands.



In labs primers don't have to be RNA, they can be chemically synthesized DNA.

Note position and orientation of primers.

DNA polymerization - Answers Taq pol + dNTPs @ 70C to create, polymerize new strands.



Taq polymerizes 5' --> 3' and requires a 3'- OH (from oligonucleotide primer).

1 full cycle of PCR - Answers You end up with 2-double stranded DNA molecules.

Thermocycler - Answers Is a PCR machine which allows amplification to proceed through successive
rounds. It raises and lowers the temperature. In order to denature, anneal, and polymerize.



Can start with small # of templates and amplify until you have more than a million molecules of DNA.
(2^40)l

To calculate amplification - Answers 2^n= double-stranded DNA segments (where n=# of cycles).

Clicker question 1: - Answers What DNA polymerase properties would you want if you could design the
perfect polymerase for PCR reactions?

A. 3' --> 5' proofreading activity --> want DNA sequence to be as accurate as possible.



C. Able to maintain activity at high temperatures --> DNA polymerase is denatured at high temps, needs
to be effective at 90C.

Taq polymerase - Answers Heat denaturation of template doesn't inactive the enzyme activity of Taq.

What makes PCR special - Answers It has been revolutionary b/c of its ease, speed, and sensitivity.



By picking primers it allows for the isolation and amplification of specific DNA fragments.

Application of PCR - Answers Detection of genetic variation for:

Forensics (DNA fingerprinting)

Medical genetic diagnostics

The benefits of buying summaries with Stuvia:

Guaranteed quality through customer reviews

Guaranteed quality through customer reviews

Stuvia customers have reviewed more than 700,000 summaries. This how you know that you are buying the best documents.

Quick and easy check-out

Quick and easy check-out

You can quickly pay through credit card or Stuvia-credit for the summaries. There is no membership needed.

Focus on what matters

Focus on what matters

Your fellow students write the study notes themselves, which is why the documents are always reliable and up-to-date. This ensures you quickly get to the core!

Frequently asked questions

What do I get when I buy this document?

You get a PDF, available immediately after your purchase. The purchased document is accessible anytime, anywhere and indefinitely through your profile.

Satisfaction guarantee: how does it work?

Our satisfaction guarantee ensures that you always find a study document that suits you well. You fill out a form, and our customer service team takes care of the rest.

Who am I buying these notes from?

Stuvia is a marketplace, so you are not buying this document from us, but from seller TutorJosh. Stuvia facilitates payment to the seller.

Will I be stuck with a subscription?

No, you only buy these notes for $7.99. You're not tied to anything after your purchase.

Can Stuvia be trusted?

4.6 stars on Google & Trustpilot (+1000 reviews)

84866 documents were sold in the last 30 days

Founded in 2010, the go-to place to buy study notes for 14 years now

Start selling
$7.99
  • (0)
  Add to cart