Question

How would you conduct an experiment using Chip-Seq technique and specific antibodies against methylated DNA, to find imprinted genes in the human genome? (Hint: start with making separate genomic DNA preparations from male and female cells).

Answer 1

To conduct a Chip-seq experiment

1. First isolate the genomic DNA here genomic DNA means DNA and histone proteins so,before isolating DNA fix the Histone proteins with DNA with the help of formaldehyde (fixative).

2. After fixation separate the genomic DNA.

3. Now shear the DNA into fragments with the help of digesting enzyme.

4. Add antibody against methylated DNA, it will bind to the place where DNA is methylated.

5. Now separate the antibody bound DNA from rest of the fragments by avidin biotin conjugation system.

6. Now separate the antibody from DNA and analyze the DNA sequence.