User:Kristina M Holton/Ovarian Notebook 2007 7 2

miRNA Extraction using 1st protocol and Qiagen miRNEasy Kit
 * Large Fibroma -> very very fatty
 * Used 3ml TRIzol from Invitrogen
 * While standing, TRIzol and fat start to pellet at the bottom; removed supernatants and centrifuged both pellet and supernatant
 * After centrifugation, already appeared to be phase separation
 * Still added glycogen + chloroform, stand RT for 10min then centrifuge 10000g 20min
 * Added EtOH: large amount of white, "SDS-like" precipitate
 * Very difficult to add precipitate to RNeasy Mini spin columns, even with additional EtOH

Differences in 2nd protocol (probable fixes):

If: no debris accumulates and the QIAzol remains pink Else: (Brown QIAzol or debris accumulation)
 * Use 2ml QIAzol for lysis
 * Skip first centrifugation step + removing the supernatant
 * Centrifuge and remove supernatant
 * Try to add all precipitate from EtOH step