Polymerase chain reaction(PCR) is a technique used in molecular biology to amplify single copy or a few copies of a piece of DNA. Step 1: Denature DNA At 95C, the DNA is denatured (i.e. The two strands are separated) Step 2: Primers Anneal At 40C- 65C, the primers anneal (or bind to) their complementary sequences on the single strands of DNA Step 3: DNA polymerase Extends the DNA chain At 72C, DNA polymerase extends the DNA chain by adding nucleotides to the 3’ ends of the primers.

2. A PRESENTATION INVERSE PCR
PRESENTED BY
1. Asraful Islam Rayhan
Dept. of Pharmacy
Jessore University Of Science &
Technology..

3. POLYMERASE CHAIN REACTION

4. Polymerase chain reaction(PCR) is a technique
used in molecular biology to amplify single
copy or a few copies of a piece of DNA.

5. Polymerase chain reaction

6. Steps of PCR
STEP 1: DENATURE DNA
At 95C, the DNA is denatured (i.e. The two strands are separated)
STEP 2: PRIMERS ANNEAL
At 40C- 65C, the primers anneal (or bind to) their complementary sequences on the single
strands of DNA
STEP 3: DNA POLYMERASE EXTENDS THE DNA CHAIN
At 72C, DNA polymerase extends the DNA chain by adding nucleotides to the 3’ ends of the
primers.

7. Denature (heat to
95oC)
Lower temperature to 56oC
Anneal with primers
Increase temperature to
72oC DNA polymerase +
dNTPs

8. Classification Of Polymerase Chain
Reaction

9. Polymerase Chain Reaction (PCRN)
Anchor
ed
Nested
Multiple
x
Reverse
transcriptase
Asymmetri
c
Quantitative Inverse RAPD Assembly

10. Applications of PCR

11. Applications of PCR
• Classification of organisms
• Genotyping
• Molecular archaeology
• Mutagenesis
• Mutation detection
• Sequencing
• Cancer research

12. Applications of PCR
• Detection of pathogens
• DNA fingerprinting
• Drug discovery
• Genetic matching
• Genetic engineering
• Pre-natal diagnosis

13. Inverse Polymerase chain reaction (iPCR)

14. Amplification of DNA of unknown sequence carried
out from known sequence.

16. Requirements for inverse PCR

17. Requirements for inverse PCR

20. Use of inverse PCR

21. Use of inverse PCR

22. Applications of inverse PCR

23. Applications of inverse PCR
Basic
Research
• Mutation screening
• Drug discovery
• Genotyping
• Molecular Epidemiology
• Molecular Ecology
• Bioinformatics
• Genomic cloning
• Site-directed mutagenesis

24. Applications of inverse PCR
Applied Research
• Genetic matching
• Detection of pathogens
• Pre-natal diagnosis
• DNA fingerprinting
• Gene therapy