User:Karmella Haynes/Notebook/PcTF Genomics/2013/01/25

01/25/13

• Illumina ChIP-seq prep: input DNA

Illumina ChIP-seq prep
> Follow manufacturer's protocol, with some modifications
> Sample (dated 12/04/10):

1. "29, FTrx fx Input" (plain cells) -- this sample was used for the original ChIP-PCR experiments (http://openwetware.org/wiki/User:Karmella_Haynes/Notebook/Polycomb_project/2011/04/22)

Perform End Repair
Note: forgot to dilute Klenow 1:5 with water before use

--> Aliquot 20 μL master mix, add 30 μL ChIP DNA
--> 20°C/ 30 min. (temp block)
--> QIAquick PCR Purification, elute with 34 μL EB

Add 'A' Bases to the 3' End of the DNA Fragments

--> Aliquot 16 μL master mix into DNA
--> 37°C/ 30 min. (heat block)
--> Zymo clean and concentrator, elute with 10 μL st.dH₂O

Ligate Adapters to DNA Fragments
Note: Did not dilute the adapter oligo mix

--> Aliquot 20 μL to DNA samples
--> 15 min./ r.t.
--> Zymo clean and concentrator, elute with 10 μL st.dH₂O
--> Store at 4°C o/n