Intro
What...?
Available
Aim
Clone Example
E and B sites
Transformation
Ligation products
pGT4 in detail
Recomb in detail
Recombs Selection
Minipreps
Gel
Blot 1
Blotting
Labeling
Blot 2
Label Detection
Blot 3
Blot and gel
AE1
AE1 (cont.)
AE2
AE2 (cont.)
PCR

Transformation

The bacteria take up the DNA very inefficiently.
And not more than one molecule per bacterium is taken up, if any..

We will analyse individual colonies formed by the bacteria on an agar plate containing the antibiotic ampicillin, to select for presence of a pGT4(-derived) plasmid..
The agar plate may look like this, after overnight growth:

Each colony represents a single clone,
harbouring a plasmid originating from one DNA molecule from the ligation reaction mixture.

The bacteria are transformed by the uptake of the DNA.
We hope to find recombinants among those transformants..

The Simple Cloning Lab          Cloning Course