Polymerase Chain Reaction WORKSHOP (3)

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Presentation transcript:

Polymerase Chain Reaction WORKSHOP (3) Presented by: Afsaneh Bazgir (Msc Student of Human Genetics) the.ocean116@gmail.com Golestan University of Medical Sciences (GOUMS)

Some types of PCR SSP-PCR Multiplex PCR Real time

Single Specific Primer PCR (SSP-PCR) There is a developed method that allows the amplification of double- stranded DNA even when the sequence information is available at one end only. This method, the single specific primer-PCR (SSP-PCR), permits amplification of genes for which only a partial sequence information is available.

Homozygote Heterozygote Homozygote

Multiplex PCR A modification of polymerase chain reaction in order to rapidly detect deletions or duplications in a large gene. This process amplifies genomic DNA samples using multiple primers and a temperature-mediated DNA polymerase in a thermal cycler. Multiplex-PCR was first described in 1988 as a method to detect deletions in the dystrophin gene.It has also been used with the steroid sulfatase gene. In 2008, multiplex-PCR was used for analysis of microsatellites and SNPs.

Applications Cont… 1. Pathogen Identification 2. High Throughput SNP Genotyping 3. Mutation Analysis 4. Gene Deletion Analysis 5. Linkage Analysis 6. Forensic Studies

Cont… With M-PCR Considerable time and effort can be saved by simultaneously amplifying multiple sequences in a single reaction. methods must be available for the analysis of each individual amplification product from the mixture of all the products. M-PCR is becoming a rapid and convenient screening assay in both the clinical and the research laboratory.

Real-time PCR Cells in all organisms regulate gene expression by turnover of gene transcripts (mRNA). The amount of an expressed gene in a cell can be measured by the number of copies of an mRNA transcript of that gene present in a sample. In order to detect and quantify gene expression from small amounts of RNA, amplification of the gene transcript is necessary. for mRNA-based PCR the RNA sample is first reverse-transcribed to cDNA with reverse transcriptase.

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