Introduction to Primer Design and PCR

About Course

Polymerase chain reaction (PCR) is a technique used to make millions of copies of a specific small amount of DNA in a laboratory. The technique was introduced in 1983 by an American biochemist named Karry Mullis, and for this valuable work he received the Nobel Prize in Chemistry in 1993.

In this course, you will learn the basics of primer design, both manually and using the appropriate software. This course is presented in such a way that even if you do not have sufficient knowledge about it, after participating in this course and a little practice, you will be able to design your primers with sufficient mastery, use PCR for various purposes, and have the ability to debug PCR.

Course Content

Introduction to PCR

Needed Equipment and preparation in the laboratory

Reaction mixture components

Properties (length, GC content, Tm …) of primers

Melting temperature, calculation method and related software

Secondary structures of primers, primer dimer and test method

Manual primer design using software under Gene Runner

How to obtain the desired DNA sequence from the NCBI

Specialized review of the existence of designed primers

Primer design with Primer 3 web application

Order primer synthesis, receive primers and prepare them for PCR

Preparing dNTP for PCR

Features DNA polymerases

Steps of performing PCR and setting the program of Thermal cycler device

Steps of horizontal electrophoresis and observation of PCR product on gel

PCR troubleshooting

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