No DNA after PCR purification with QiaQuick?
Posted 09 May 2011 - 06:23 PM
I'm having a curious problem, and was wondering if you might be able to help.
I'm amplifying out 3' UTRs in several genes of interest, and I've gotten decent yields. Gels look beautiful; clear, defined bands exactly where I want them. My only problem is, when I try to clean up the reaction with a kit (QiaQuick), I lose all my product.
I dug through the forum and tried the various tips (checked pH of water; tried using elution buffer instead; heated it up and let it sit for 10 minutes or so before eluting; etc), but after eluting and NanoDrop-ing, there's nothing. I'm going to try and do alcohol precipitation tomorrow, but I was wondering if you might have any tips on what could be happening.
Posted 10 May 2011 - 03:53 AM
Are you using the QBI buffer which has the indicator? The indicator assures that the pH of the dissolved gel is correct for column binding.
Posted 10 May 2011 - 06:59 AM
Posted 10 May 2011 - 04:33 PM
...but after eluting and NanoDrop-ing, there's nothing.
What does a cleaned-up sample look like on a gel?
Posted 14 May 2011 - 07:50 AM