Purification of RNA from Cells and Tissues by Acid Phenol-Guanidinium Thiocyanate-Chloroform Extract Single-step technique, cells are homogenized in guanidnium thiocyanate and the RNA is purified from the lysate by extraction with phenol:chloroform at reduced pH. Many samples can be processed simultaneously and speedily. The yield of total RNA depends on the tissue or cell source and is generally in the range of 4-7 µg/ml starting tissue or 5-10 µg/106 cells. IMPORTANT: Prepare all reagents used in this protocol with Diethyl pyrocarbonate (DEPC)-treated H2O.
Removal of Genomic DNA from Total RNA for Use in Fluorescent mRNA Differential Display Protocol Protocol is the second in a set of three, describing fluorescent mRNA differential display (FDD or FDDRT-PCR). For the purposes of FDD gene expression analysis, as well as any other RNA-based gene expression technologies, contaminating genomic DNA must be removed before reverse transcription and subsequent PCR.
RNA Preparation from Cultured Cells or Tissue Samples Protocol Protocol for RNA preparation from cultured cells or tissue samples. This protocol has been used to isolate RNA from relatively small tissue samples. The RNA is clean enough for Rnase protection, cDNA synthesis, and RT-PCR analysis.
