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प्रश्न
What is PCR? Explain different steps involved in it.
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उत्तर
Definition: Polymerase Chain Reaction (PCR) is the process of in vitro amplification of gene of interest using a PCR machine.
Mechanism of PCR: At the start of PCR, all the requirements are mixed together in ‘eppendorf tube’ and the following operations are performed sequentially:
Step i: Denaturation
The reaction mixture is heated to a temperature (90–98o C) to separate two strands of desired DNA. This is called denaturation.
Step ii: Annealing
The mixture is allowed to cool (40–60o C) that permits pairing of the primer to the complementary sequences in DNA. This step is called annealing.
Step iii: Primer extension / Polymerization
The temperature (70–75o C) allows thermostable Taq DNA polymerase to use single-stranded DNA as template and adds nucleotides. This is called primer extension. It takes around two minutes duration.
संबंधित प्रश्न
State the role of DNA ligase in biotechnology.
Which of the following device is required for PCR?
Match the Column I (Restriction enzyme) with Column II (Recognition sequence) and select the correct option.
| column I (Restriction Enzyme) | column II (Recognition sequence) | ||
| i. | Alu | P. | 5'---G- ↓-G-A-T-C-C---3' 3'---C-C-T-A-G- -G---5 |
| ii. | BamHI | Q. | 5'--- A-G- ↓ -C-T---3' 3' ---T-C- G-A--- 5' |
| iii. | Eco RI | R. | 5'--G-T-C- ↓ -G-A-C--3' I 3'--C-A-G-t-C-T-G--5' |
| iv. | HindII | S. | 5'---G- ↓ -A-A-T-T-C---3' 3'---C-T-T-A-A- -G---5' |
The following identifies the major features of technology that differentiate modem biotechnology from classical biotechnology.
i. Capability of science to change the genetic material for getting new specific products through rDNA technology, polymerase chain reaction (PCR), microarrays, cell culture and fusion, and bioprocessing.
ii. Ownership of technology and its sociopolitical impact.
iii. Modem biotechnology makes use of only fermentation technology.
iv. Classical biotechnology relies only on principles of plant and animal tissue culture.
In PCR technique, the organism providing thermostable DNA polymerase is a ____________.
Lytic cycle involves bacteriophage and ____________.
What would happen when one grows a recombinant bacterium in a bioreactor but forget to add antibiotic to the medium in which the recombinant is growing?
Describe three steps involved in mechanism of PCR.
What are the different types of electrophoresis?
Which one of the following enzymes is used to join DNA fragments?
