If a desired gene is identified in an organism for some experiments, e

1.	If a desired gene is identified in an organism for some experiments, explain the process of the following : (i) Cutting this desired gene at specific location. (ii) Synthesis of multiple copies of this desired gene.
Answer» (i) - Identifying the restriction endonuclease that recognises the palindromic nucleotide sequence of the desired gene. - The restriction endonuclease inspects the DNA sequences - finds and recognises the site - Cuts each of the double helix at the specific point - A little away from the centre of the palindromic site - between the same two bases on the opposite strand. - Makes the over hanging stretch single stranded portion as a sticky end. (ii) - By PCR/Polymelase Chain Reaction - Desired gene is synthesised in vitro - DNA is denatured - Annealed using two sets of primers - Thermostable Taq polymerase extends the primers using nucleotides (provided in the reaction and genomic DNA as template) - Amplified fragments are ligated.

If a desired gene is identified in an organism for some experiments, explain the process of the following : (i) Cutting this desired gene at specific location. (ii) Synthesis of multiple copies of this desired gene.

Answer»

(i) - Identifying the restriction endonuclease that recognises the palindromic nucleotide sequence of the desired gene.

- The restriction endonuclease inspects the DNA sequences

- finds and recognises the site

- Cuts each of the double helix at the specific point

- A little away from the centre of the palindromic site

- between the same two bases on the opposite strand.

- Makes the over hanging stretch single stranded portion as a sticky end.

(ii) - By PCR/Polymelase Chain Reaction

- Desired gene is synthesised in vitro

- DNA is denatured

- Annealed using two sets of primers

- Thermostable Taq polymerase extends the primers using nucleotides (provided in the reaction and genomic DNA as template)

- Amplified fragments are ligated.