Posted 21 July 2010 - 02:19 PM
I did a stupid mistake today and am thinking what to do except waiting for the gel pic what i will get after 2 days.
I use phenol chloroform to extract DNA from cells. So i treat the cells with extraction buffer and it has rnase a. After that i treat with proteinase K and leave overnight. But today instead of adding proteinase K, i added Rnase A again and now conc of rnase a is really really high in my sample. I talked to Sigma about the inactivation temp of rnase a, but they dont know.
So my question is, how can high conc of rnase a affect ds DNA provided Rnase A is Dnase free?
Any help would be appreciated.
Posted 21 July 2010 - 04:56 PM
Provided the RNAse is DNAse free, excess RNAse will not have any effect on your DNA sample. You might want to remove this RNAse if you are going to transform/transfect your DNA sample into cells.
Posted 23 July 2010 - 03:34 AM