Protocol Online logo
Top : New Forum Archives (2009-): : ChIP and Next Generation Sequencing

chip precipitation - using isopropanol in chip precipitation (Jul/26/2011 )

At the end of PK step on chip experiment and as I was using 1.5 ml eppendorf and my volume of eluted was 550 ul I used isopropanol 0.8 vol with 1/10 AcONa 3M and glycogen, I put in the freeze -80, centrifuge but I could not see any pellet. I let the tubes at -20C o/n. question: It was a bad idea to use isopropanol, I need more alcohol volume to precipitate the DNA, Waht could I do to repare the missfit?.
Thank you.


sounds to me like you may have skipped a cleaning up the PK reaction with a phenol:chloroform:isoamyl extraction, then go on to precipitate your DNA with EtOH. Or even simpler, after your PK reaction, use a Qiagen column.