Help: RNaseA or Proteinase K or both for DNA extraction in CHIP - (Nov/09/2007 )
I am using Upstate CHIP kit (Catalog # 17-295).
The kit protocol says to do the following with the chromatin eluate
1 Reverse Cross link
2 Proteinase K digestion
3 phenol/chloroform extraction and ethanol precipitation
However, in step3, I plan to use Qiaquick PCR purification kit instead.
Could anyone kindly tell me how you treat the eluate before processing it using the PCR kit? For example:
Do I use RNase A or Proteinse K or both?
how long? temperature? Concentration?
Together or in separate steps?
Many thanks in advance!
Do proteinase K digestion first, and then RNase treatment. Just follow the protocol for amount, temperature and duration. the purify use qiaquick spin column by adding 5 volume of PB buffer. You need to buy extra PB buffer or prepare your only because the volume needed is big.
I will try that tomorrow! Thanks a lot, pcrman!
What is the reason for swapping the proteinase k/rnase steps?