Workshop on PCR

Last Date to Register - 30th August, 2016.

To Register online click on this link: https://docs.google.com/forms/d/e/1FAIpQLSf8vcWlbx_ZWCvNPJzn9eDdoLV_UfR1QPHPvrS-sOSf99MDnA/viewform?c=0&w=1

To download the offline Registration Form, click here

Polymerase Chain Reaction

Polymerase chain reaction (PCR) is a molecular technique to amplify a particular section of DNA. It was developed in 1983 by the American biochemist Kary Mullis. PCR can be used in generating billions of copies of a particular section of DNA from a very low amount in lesser time.

Components of PCR mix:

Five core ingredients are required to set up a PCR. The components are:

• ·        DNA template - the gene of interest
• ·        Primers - short DNA sequence that are complementary to the template DNA and bind to either side of the section of DNA
• ·        Deoxy nucleotide triphosphates (dNTPs) - consist of dATP, dCTP, dGTP, and dTTP. These nucleotide bases A, T, G, and C, are essentially "building blocks" used for the synthesis of new DNA strands.
• ·        Taq DNA polymerase enzyme - a thermo-tolerant DNA polymerase from Thermus aquaticus, a species of bacterium found in hot springs which add DNA bases complementary to the template.
• ·        PCR buffer - contains KCl, MgCl₂, BSA and Tris base to create the environment conducive for PCR to take place.

PCR Tube Containing components

PCR tubes

 PCR involves thermal cycling, a process of heating and cooling which is carried out by thermocycler.

Thermocycler holding the PCR tubes

                          

Steps involved in PCR:

Denaturation, annealing, and extension are the three major steps involved in PCR. These three steps form a cycle and are repeated 35 times, doubling the number of DNA copies after each cycle. A result is a huge number of copies of the specific DNA segment produced in a relatively short period.  The copies of DNA are called as amplicon. The three steps are:

1.  Denaturation: During this step, the PCR reaction mix containing the double-stranded DNA template is heated to 95⁰C for 30 seconds.  The high temperature causes the hydrogen bonds between the bases in the two strands of DNA to separate it into two single strands. These two single strands of DNA act as templates for the synthesis of the new strands of DNA.
2.  Annealing: During this step, the reaction mix is cooled to 55⁰C for 30 seconds. This enables the primers to attach to a specific location on the single-stranded template DNA by way of hydrogen bonding. Primers serve as the starting point for DNA synthesis.
3.  Extension: During this step, the temperature is increased to 72⁰C for 1 minute to enable the new DNA strand to be made by Taq DNA polymerase enzyme. The optimum temperature for the Taq polymerase to build the complementary strand is 72⁰C. The enzyme attaches to the primer and then adds DNA bases to the single strand one-by-one in the 5’ to 3’ direction. The result is a new strand of DNA and a double-stranded molecule of DNA. The duration of this step depends on the length of DNA sequence being amplified but usually takes around one minute to copy 1,000 DNA bases (1Kb).

Mechanism of PCR

             

              The amplicon will be checked on 1% agarose gel containing ethidium bromide (EtBr). The EtBr which is added to the gel intercalates between the nucleotide bases and makes the amplicon to fluoresce upon illumination with ultraviolet light. The amplicon can be visualized by illuminating the gel under the ultra violet light. The size of the amplicon can be known by running marker along with the sample. 

Agarose gel photograph of the amplicon run  along with the marker in 1% gel

                

Applications:

PCR is widely used in medical and biological research labs. It is used for:

·               Medical applications like genetic testing, to check the presence of mutations

·     Infectious disease applications to detect the presence or absence of a gene for identifying pathogens during infection

·       Forensic applications for generating forensic DNA profiles from tiny samples of DNA, and for DNA paternity testing

·              Research applications like sequencing, genetic mapping, gene expression and many others.

Venue: Department of Genetic Engineering,

School of Bioengineering, 

SRM University, 

Kattankulathur, Chennai-603203

Workshop Date: September 6, 2016

Workshop Timing: 9.00 a.m. to 5.00 p.m.

Registration Details: The registration fee for the workshop is Rs.200/- per participant.

Registration fee includes:

·       Participation in the workshop

·       Refreshment and lunch

·       Workshop manual

·       Certificate for successful completion

 Note:  

1. Transportation is not included in the registration fee. Participants will have to arrange for their own transport.
2. No accomodation will be provided for outstation students.
3. Atleast one teacher or parent should accompany the students

Eligibility: 12^th standard school students, 5 per school 

Mode of payment: DD/Cheque/online transfer

DD should be taken in favour of “Genetic Engineering Association”, payable at Chennai 

     Cheque should be in the name of “Genetic Engineering Association” 

     DD/ Cheque should be sent to the following address on or before 30^th August 2016

The Head of the Department,

Department of Genetic Engineering

School of Bioengineering,

SRM University,

Kattankulathur, Chennai 603203.

  

Online Transfer (NEFT): Bank Name: City Union Bank

                                            Branch: Tambaram   

                                            Account Name: Genetic Engineering Association    

                                            Account No: 117001000403689    

                                            IFSC code: CIUB0000117

For further details contact: Ms. S. Poovitha.

Mobile No. 9884604011

Last date to Register - 30th August, 2016.

To Register kindly click on this link: https://docs.google.com/forms/d/e/1FAIpQLSf8vcWlbx_ZWCvNPJzn9eDdoLV_UfR1QPHPvrS-sOSf99MDnA/viewform?c=0&w=1

Total no. of Seats: 100

  

To download the offline Registration Form, click here