Biotechnology: Principles and Processes — Part 2: Processes of recombinant DNA technology: visual lesson, CBSE Class 12 Biology

Lesson

Processes of recombinant DNA technology

Part 2 — the steps from extracting DNA to making a finished product.

In this lesson you will

List the steps of recombinant DNA technology in order
Explain DNA isolation, cutting, PCR amplification and ligation
Describe transformation, selection and growth in bioreactors
Explain downstream processing

Concept

Step 1: isolating DNA

DNA is enclosed in membranes, so the cell must be broken open. Then other macromolecules are removed, and DNA is precipitated.

Table of treatments used to isolate pure DNA
Problem	Treatment
Bacterial cell wall	Lysozyme
Plant cell wall	Cellulase
Fungal cell wall	Chitinase
RNA	Ribonuclease
Proteins (histones)	Protease
Getting DNA out of solution	Add chilled ethanol: DNA precipitates as fine threads (spooling)

Concept

Step 2: cutting DNA

Purified DNA is incubated with a restriction enzyme under optimal conditions. Agarose gel electrophoresis checks the progress of digestion. The vector is cut with the same enzyme, and the gene of interest is joined to it.

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