Question

1. Restriction endonuclease, DNA Ligase.
2. Sticky ends, blunt ends.
3. Kary Mullis in 1983 received Nobel Price for development of PCR technique.
4. PV 92 gene, homozygous, heterozygous.

Answer 1

POLYMERASE CHAIN REACTION (PCR):-

# PCR stand for polymerase chain reaction.

#PCR technique was invented by KARRY MULLIS

#Each cycle has three steps (i) Denaturation

(ii)Primer annealing (iii) Extension of primers #Thermal cycle is used in PCR for Denaturation melting. Denaturation of DNA at 70 Celsius is due to breakdown of hydrogen bond .

#In PCR multiple copies of gene of interest is synthesized in vitro using two set of primers and the enzyme DNA polymerase. Primer provide 5' site to the enzyme the enzyme extends the primer using the nucleotide provided in the reaction and the genomic DNA as template.

#if the process of replication of DNA is repeated many times the segment of DNA can be amplified to approximately billion times.

#Such repeated amplification is achieved by the use of thermostable DNA polymerase which is isolated from a thermophilic bacterium thermus aquaticus which remain active during the high temperature induced denaturation of double stranded DNA polymerase is used between annealing and extension step of PCR .

#Amplified fragment of DNA can now be used to like it with the help of further cloning.