2. Polymerase Chain Reaction WORKSHOP (3)
Presented by: Afsaneh Bazgir
(Msc Student of Human Genetics)
Golestan University of Medical Sciences
(GOUMS)

3. Some types of PCR
SSP-PCR
Multiplex PCR
Real time

4. 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.

5. Homozygote Heterozygote Homozygote

6. 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.

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

8. 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.

10. 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.

11. THANK YOU