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

Cut and Paste

Digest the vector and donor DNAs with the same restriction enzymes.
  • E= EcoRI
  • B=BamHI

Mix all DNA fragments and add the enzyme ligase.

= "cut and paste"

Result: many combinations of DNA molecules possible.

After adding CaCl2 to an E.coli bacterial culture, to enable the bacteria to take up foreign DNA, we will let these so-called competent cells  take up the DNA molecules from the ligation reaction mixture.

The Simple Cloning Lab          Cloning Course