Catalog #: 10000 (Total RNA purification from tissue/cells; 10001 (mRNA purification from tissue/cells); 10002 (mRNA purification from total RNA)
The production of a high quality library containing a large number of full-length cDNA clones requires that the tissue and RNA preparation be performed in a certain way. These methods will help to guarantee that the starting material and ultimately the cDNA library are not compromised.
To isolate total RNA from frozen tissue, the tissue is ground to a fine powder using liquid nitrogen and a mortar and a pestle. To isolate RNA from cells, the cells are pelleted and the pellet washed with PBS. Total RNA is quickly isolated by homogenization or lysis using a reagent, such as TRIzol or equivalent. Total RNA is further processed to mRNA using poly A+ selection. Total RNA and mRNA are dissolved in nuclease-free water (no DEPC). We provide quality controlled RNA from customer supplied materials.
The A260/A280 ratio of the RNA must be at least 1.8.
Gel analysis of the total RNA must show that the 28S rRNA band is nearly twice the intensity of the 18S rRNA band.