Class 12 Zoology simulations · Biotechnology: Principles and Processes

Making recombinant DNA

Cut a plasmid and foreign DNA with the same restriction enzyme, join them, transform bacteria and pick out the recombinants.

Open the simulation

What to notice

An insert inside a selectable-marker gene inactivates it — insertional inactivation reveals which colonies are recombinant.

Key relation

EcoRI site: 5′-GAATTC-3′ (palindrome) → sticky ends

Try this

Choose blue–white screening. What colour are recombinant colonies?

What you can change

• Step: 1 to 4
• Screening method: choose Blue–white (lacZ, β-galactosidase), Insertional inactivation of tetᴿ (pBR322)

More in Biotechnology: Principles and Processes

• Agarose gel electrophoresis
• Polymerase chain reaction (PCR)

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