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Q.Mention processes of Recombinant DNA Technology.

Uttarakhand UbseUttarakhand Board Intermediate (Class 12) 2019Subjective· 3mImportance★★★★★
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Steps of rDNA technology: isolate DNA → cut with restriction enzymes (isolate gene) → amplify (PCR) → ligate gene into a vector (ligase) → transform into a host cell → culture & select transformants → recover the product (downstream processing).

Concept

Recombinant DNA (rDNA) technology / genetic engineering is the set of techniques used to isolate a desired gene, join it to a vector, and introduce it into a host organism so that the host expresses the gene and produces a desired product.

Main processes (steps)

  1. Isolation of the genetic material (DNA): the DNA is released from the cells by breaking them open (using enzymes like lysozyme, cellulase) and is purified (removing proteins, RNA, etc.), then precipitated with chilled ethanol.
  2. Cutting/fragmentation of DNA (restriction digestion): the DNA is cut at specific recognition sites using restriction endonucleases ("molecular scissors"), producing fragments with sticky ends; the same enzyme cuts both the source DNA and the vector so their ends are complementary. The desired gene is separated by gel electrophoresis.
  3. Amplification of the gene of interest: the required gene may be amplified using PCR (Polymerase Chain Reaction) with primers and Taq polymerase.
  4. Ligation into a vector (making recombinant DNA): the gene fragment is joined to a cloning vector (e.g. a plasmid) using the enzyme DNA ligase, forming the recombinant DNA molecule. …

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