Found 2 discussions for this experiment
1 year ago
1 year ago by Paul G. Macon
How do I extract RNA from animal tissue without using liquid nitrogen? I tried the RNA extraction by using the TRIzol reagent and I homogenize the tissue using polytron homogenizer at room temperature for 30secs is this correct?
1 year ago
1 year ago by Aaron Stege
I used a serum/plasma kit for my serum samples. After the phase separation the samples should have 3 phases: a colourless aqueous phase, a white interphase and a red organic phase. However, in some of my samples there was no aqueous phase unless I wait for an extended period of time. How can I circumvent this problem?
Found 1 matching solution for this experiment
|We employed the same method for RNA isolation for these different tissues as was used for our previously-published brain RNA isolation study, using the RNA EasyPlus Mini Kit (Qiagen, Valencia, CA) (Birdsill et al. 2011). Each tissue sample was dispersed in RNA-lysis buffer (containing 0.1 M β-mercaptoethanol) by mild sonication (10–15 s), and then processed according to the manufacturer’s instructions. In brief, extracted tissue is passed through a prefilter to remove DNA, adjusted with an equal volume of 70 % ethanol and applied to the RNA column. After washes with kit buffers, RNA is eluted from the column with 100 μl of water. A point to note was that for highest yield and integrity, sonication of tissue sample should be carried out for as brief a time as necessary to disrupt the tissue; excessive sonication significantly reduced the yield of RNA from tissues. Some tissues contained insoluble connective tissue or fat that did not dissolve in RNA lysis buffer, but were removed on the DNA prefilters prior to RNA isolation. This same method was used for all cerebellum brain samples.|