Detailed polymerase chain reaction concepts

Description

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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