isopropanol precipitate lysis? - (Sep/11/2005 )
when extracting HIV RNA from plasma samples, we use different kits. One of them (which seems to be giving the best results with subsequent RT-PCR) is very expensive and every buffer and so on is patented. We know the principle it relies on (just spin down virus from plasm for 1 hour and lyse the pellet after which isopropanol precipitation is performed). To make things cheaper, we want to try to do this with our own lysis buffer (or lysis buffers that are available from f.i. qiagen). We are not 100% sure if lysis buffers contain enough ions for isopropanol precipitation to be effecient.
So, can anyone explain the exact need (pH, concentration of ions) for efficient isopropanol (or ethanol, even though this one is claimed to be less efficient by the companies) precipitation? Or anyone with experience on precipitating a lysis solution?
If the buffers are patented you can most likely find out how to make them by examining the patent descriptions. Go to www.uspto.gov and search for the patent number, which should be somewhere in the literature that came with the product.
Hope that helps,