Plasmid miniprep w/ Glass fiber filter plates ????
Posted 17 August 2010 - 08:19 AM
To do this we are following the protocols set out by both Millipore (http://www.millipore...ons/tech1/tn004) and Corning (http://catalog2.corn...alsp_an_017.pdf).
Both of these protocols use a method that involves 6M KI solution as a "bind solution". However this is causing real problems as it is impossible to read on a spectro/nanodrop. (huge peak at 250nm) and eluted DNA just floats out of the wells on an agarose gel (no matter how many washed with 80% Ethanol).
When we carried out a test with our plates using protocol or Qiagen solutions (without KI) we avoided the problem but the yield was very very low (<10ng/ul).
However, when we used Qiagen PB buffer (binding) as the "optional wash step" the yield went up to ~30-40ng/ul (still not excellent but an improvement). I believe Buffer PB does not contain potassium iodide (KI).
My questions are:
Is there any way to avoid the KI bind solution? What can we use instead?
Are glass fiber plates the best plates for this task?
Or is there another protocol that anyone knows of for Glass fiber plates that uses a different bind solution?
Posted 17 August 2010 - 10:42 AM
Posted 17 August 2010 - 06:08 PM