An improved method for the extraction of total RNA from ribonucleases rich tissues
2006
Abbas, N. (Punjab Univ., Quaid-e-Azam Campus, Lahore (Pakistan). School of Biological Sciences) | Abbas, Z. (Punjab Univ., Quaid-e-Azam Campus, Lahore (Pakistan). School of Biological Sciences) | Akhtar, M. (Punjab Univ., Quaid-e-Azam Campus, Lahore (Pakistan). School of Biological Sciences)
The ability to isolate clean, intact RNA has important uses in cloning genes and is essential to analyze gene expression. To get useful clones, it is critical that full-length RNA is used as the starting material. To obtain good quality of eukaryotic mRNA it is necessary to minimize the activity of ribonucleases liberated during cell lysis by using their inhibitors. Guanidinium thiocyanate and 2-mercaptoethanol containing buffer ensures thorough denaturing of macromolecules and inactivation of RNases. Many different protocols and commercial kits are available for isolation of RNA from tissue but all have their own limitations. A single step isolation method for RNA was modified to get good quality total RNA from RNases rich tissue e.g., pancreas. Tissue samples (chick and bovine pancreas) were collected under RNases free conditions. Total RNA was extracted using guanidinium thiocyanate and acid phenol.
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