Detailed polymerase chain reaction concepts

Part 1:

what is PCR?

who invented PCR?

How does PCR works?

PCR  chemical ingredients

PCR Diagram

Steps:

1 – Denaturation

2- Annealing

3-Extension

Part 2:

Types of PCR

Quantitative

Real time

Reverse transcription

Part 3:

Multiplex

Nested

High -fidelity

fast

heart start

GC rich

Long range

AP PCR

Part 4:

application of PCR

clinical diagnosis, prenatal diagnosis, retro-viral, bacterial infection, cancer diagnosis, sex determination embryos and forensic medicine

Standard ingredients in the mixture are:

·the DNA segment of interest

·specific primers

·heat-resistant DNA polymerase enzyme

·the four different types of DNA nucleotides

·the salts needed to create a suitable environment for the enzyme to act.

Quantitative PCR (qPCR), also called real-time PCR or quantitative real-time PCR, is a PCR-based technique that couples amplification of a target DNA sequence with quantification of the concentration of that DNA species in the reaction.

Real–time PCR is the technique of collecting data throughout the PCR process as it occurs, thus combining amplification and detection into a single step. This is achieved using a variety of different fluorescent chemistries that correlate PCR product concentration to fluorescence intensity

Reverse transcription polymerase chain reaction is a laboratory technique combining reverse transcription of RNA into DNA and amplification of specific DNA targets using polymerase chain reaction. It is primarily used to MEA

sure the amount of a specific RNA.

Direct detection of microorganisms in patient specimens

Identification of microorganisms grown in culture

Detection of antimicrobial resistance

Investigation of strain relatedness of a pathogen of interest

Genetic fingerprinting (forensic application/paternity testing)

Detection of mutation ( investigation of genetic diseases)

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