VPB 321: Animal Biotechnology (2+1)

TYPES OF PCR TECHNIQUES

• There are several types of PCR used. The below mentioned are the most commonly used types of PCR.

RT-PCR

• Reverse transcriptase polymerase chain reaction (RT-PCR) is used to convert the RNA template into complementary DNA (cDNA) by using the enzyme reverse transcriptase and then the target is amplified into many copies through PCR steps. RT-PCR is applied to diagnose RNA viruses, mRNA expression etc.

Nested PCR

• Nested PCR is a conventional PCR with a second round of amplification using a different set of primers. This second set of primers is specific to a sequence found within the DNA of the initial conventional PCR amplicon. The use of a second amplification step with the "nested" primer set results in a reduced background from products amplified during the initial PCR due to the nested primers’ additional specificity to the region. The amount of amplicon produced is increased as a result of the second round of amplification.

Multiplex PCR

• Multiplex PCR is the term used when more than one pair of primers is used in a PCR. The goal of multiplex PCR is to amplify several segments of target DNA simultaneously and thereby to conserve template DNA, save time, and minimize expense. It is a PCR strategy that enables the amplification of multiple DNA targets in one run.

Real time PCR

• Real-Time PCR, also called quantitative (real-time) PCR or Real-Time Quantitative PCR (RTQ-PCR), is a method of simultaneous DNA quantification and amplification.