The document discusses the history and development of the polymerase chain reaction (PCR) technique. It describes how Kary Mullis invented PCR in 1985 and was awarded the Nobel Prize for it. It then explains the basic steps of PCR including denaturation, annealing of primers, and extension. Finally, it discusses several variations and applications of PCR including real-time PCR, asymmetric PCR, and comparisons to cloning techniques.

1. POLYMERASE CHAIN REACTION POLYMERASE CHAIN REACTION “ Hands on training in Biotechnology” (2011) Centre of Excellence in Agri-Biotechnology, SVPUAT,Meerut,UP. R.Sujatha, Scientist B, New Delhi.

3. History OF PCR As is the photo copier a basic requirement in an office,so is the PCR machine in a molecular biology Laboratory !!!!!!!!! PCR is DNA replication in a test tube……..

5. DNA ?

17. NEW AUTOMATED PCR OLD PCR

33. Parameter PCR Gene cloning 1. Final result Selective amplification of specific sequence Selective amplification of specific sequence 2. Manipulation In vitro In vitro and in vivo 3. Selectivity of the specific segment from complex DNA First step Last step 4. Quantity of starting material Nanogram (ng) Microgram (m) 5. Biological reagents required DNA polymerase (Taq polymerase) Restriction enzymes, Ligase, vector. bacteria 6. Automation Yes No 7. Labour intensive No Yes 8. Error probability Less More 9. Applications More Less 10. Cost Less More 11. User’s skill Not required Required 12. Time for a typical experiment Four hours Two to four days