Polymerase Chain Reaction

50 %
50 %
Information about Polymerase Chain Reaction

Published on September 26, 2008

Author: rkduary

Source: authorstream.com

Slide 1: Polymerase Chain Reaction Raj Kumar Duary Molecular biology unit, Dairy microbiology Division, National Dairy Research Institute Karnal, India-132001 E-mail : rkduary@gmail.com Specific synthesis of DNA in vitro via a PCR reaction. (Emeryville; California, Cetus) : Specific synthesis of DNA in vitro via a PCR reaction. (Emeryville; California, Cetus) Invented by- Kary Mullis (1983) Shared the Nobel Prize in Chemistry with Michael Smith in 1993. Slide 3: PCR is a rapid, inexpensive and simple way of copying specific DNA fragments from minute quantities of source DNA material • It does not necessarily require the use of radioisotopes or toxic chemicals • It involves preparing the sample DNA and a master mix with primers, followed by detecting reaction products Slide 4: Chain reaction relies on DNA replication process Repeated cycles of melting (strand separation), primer annealing, and primer extension by cycling temperatures Need a tough enzyme to deal with high temperatures Polymerases isolated from thermophilic bacteria (Thermus aquaticus, Pyrococcus furiosus) Slide 5: ! Denaturation: DNA fragments are heated at high temperatures, which reduce the DNA double helix to single strands. These strands become accessible to primers ! Annealing: The reaction mixture is cooled down. Primers anneal to the complementary regions in the DNA template strands, and double strands are formed again between primers and complementary sequences ! Extension: The DNA polymerase synthesizes a complementary strand. The enzyme reads the opposing strand sequence and extends the primers by adding nucleotides in the order in which they can pair. The whole process is repeated over and over PCR procedures: steps Slide 6: Melting temperature Tm°C = 2(A/T) + 4(G/C) Tm oC Temperature at which half possible H bonds are formed Slide 7: The Thermus aquaticus DNA polymerase (Taq) Not permanently destroyed at 94ºC Optimal temperature is 72ºC Does not have proof reading ability Error rate 1 in 2 X 104 bases Seems rare but can be recovered in cloning a single molecule Newer polymerases have high fidelity : Does not have proof reading ability Error rate 1 in 2 X 104 bases Seems rare but can be recovered in cloning a single molecule Newer polymerases have high fidelity Problems with Taq Slide 9: 

Add a comment

Related presentations

Related pages

Polymerase-Kettenreaktion – Wikipedia

Die Polymerase-Kettenreaktion (englisch Polymerase Chain Reaction, PCR) ist eine Methode, um die Erbsubstanz DNA in vitro zu vervielfältigen. Dazu wird ...
Read more

Polymerase chain reaction - Wikipedia, the free encyclopedia

The polymerase chain reaction (PCR) is a technology in molecular biology used to amplify a single copy or a few copies of a piece of DNA across several ...
Read more

Polymerase Chain Reaction (PCR) - National Center for ...

Polymerase Chain Reaction (PCR) Introduction PCR (Polymerase Chain Reaction) is a revolutionary method developed by Kary Mullis in the 1980s. PCR is based ...
Read more

"Polymerase Chain Reaction (PCR)" Biology Animation ...

Polymerase Chain Reaction. Polymerase chain reaction (PCR) enables researchers to produce millions of copies of a specific DNA sequence in approximately ...
Read more

Polymerase Chain Reaction: Learn Definition and Steps of PCR

Learn about PCR (polymerase chain reaction) a method of analyzing a short sequence of DNA or RNA. PCR has many uses, diagnostic, forensics, cloning, and more.
Read more

Polymerase Chain Reaction (PCR) - PCR Fact Sheet

Fact sheet on polymerase chain reaction (PCR), published by the National Human Genome Research Institute.
Read more

Polymerase Chain Reaction - Plattsburgh State Faculty and ...

The Polymerase Chain Reaction. The Polymerase Chain Reaction (PCR) provides an extremely sensitive means of amplifying small quantities of DNA. The ...
Read more

Polymerase Chain Reaction (PCR) - YouTube

Polymerase chain reaction (PCR) allows researchers to amplify DNA in a test tube. This process uses an enzyme derived from heat-resistant bacteria.
Read more

Polymerase chain reaction - definition of polymerase chain ...

polymerase chain reaction n. Abbr. PCR A technique for amplifying DNA sequences in vitro by separating the DNA into two strands and incubating it with ...
Read more

PCR - Learn Genetics

PCR (short for Polymerase Chain Reaction) is a relatively simple and inexpensive tool that you can use to focus in on a segment of DNA and copy it billions ...
Read more