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Making Recombinant DNA

Manual: General · Subject: Biology

Learn how a gene is inserted into a plasmid to create recombinant DNA.

Recombinant DNA

Definition

Recombinant DNA is DNA made by joining pieces from different sources. In cloning, this usually means combining a plasmid with a gene of interest.

How recombinant DNA is made

  1. 1

    Step 1: Cut the plasmid open with a restriction enzyme.

  2. 2

    Step 2: Cut out or prepare the gene of interest.

  3. 3

    Step 3: Mix the gene and plasmid so the ends can match.

  4. 4

    Step 4: Use DNA ligase to seal the DNA together.

  5. 5

    Step 5: Check that the plasmid now contains the gene.

ℹ️

Gene of interest

The gene of interest is the DNA piece the scientist wants to study or copy.

Why this matters

Once the gene is inside the plasmid, it can be carried into a host cell and copied many times.

Before and after ligation

Before

  • Plasmid is cut open
  • Gene is separate

After

  • Gene is inserted into plasmid
  • New recombinant DNA molecule forms

What is recombinant DNA?

What is the purpose of DNA ligase in cloning?

The gene of interest is usually inserted into a: