Skip to the content.

ChIP protocol

Preparation of mycelium

Culture

Fixation

Grinding and storage (In cold room)

Day 1

Lysis and chromatin digestion

(Possibility of storing -80 ° C at this stage, but for the ChIP it is better to avoid the thawing steps).

Checking of chromatin fragmentation (nucleosomal digestion pattern)

Day 2

Phenol extraction of the DNA

Adjust the volume to 300 µl with H2O. Carefull! Phenol and chloroform must be used under the fume hood!

To the tube kept over-night at 65°C, add:

Ethanol precipitation:

It should looks like this:

ChiP:

Dilution the chromatin :

Prepare as many tubes as necessary for the different IPs and controls you will need

In the cold room:

Caution: 1 Tube (5μg) = 1 Precipitation with 1 antibody. Prepare 1 tube for each replicate of each antibody.

Day 3

Binding and washes

Elution

Day 4

DNA Purification

DNA Precipitation