Qiagen Midi Prep Question
Posted 19 December 2010 - 01:19 PM
Posted 19 December 2010 - 06:00 PM
Posted 19 December 2010 - 09:02 PM
Yes, I eluted the DNA (after washing the column with 20ml QC buffer), then added the isopropanol, also through the column, instead of directly to the eluate. I am hoping there was nothing left on the column that would have contaminated the DNA.
Posted 20 December 2010 - 05:08 AM
Posted 20 December 2010 - 06:20 PM
Hi - Yes, I thought it wouldn't be a problem, too. However, after nanodropping the samples today, I can say "Don't do what I did!". The DNA was completely degraded. My thought is that perhaps the isopropanol pulled some column material into the sample. I don't know what the columns are made of, but clearly something came off that destroyed the DNA.
Posted 20 December 2010 - 06:27 PM
Posted 21 December 2010 - 06:38 AM
Posted 21 December 2010 - 09:21 PM
The odd ratios do indicate that the DNA sample is contaminated by something... what do intend to use this DNA for?
I believe a phenol chloroform extraction should clear up most contaminants.
And yes, do run the DNA on a gel to check for integrity.
Posted 21 December 2010 - 09:56 PM