Isolation of Total RNA from Rat Liver Tissue by Improved Trizol Method

Objective To improve a technique that is employed to isolate and identify high purity and integrity of total RNA extracted from fresh animal tissue.Methods Referring to the RNA isolation method proposed by Chomczynski P(1987),we improved the protocol of RNA isolation.Furthermore,we adjusted the conventional agarose gel electrophoresis to be better used to identify the RNA quality.Results When total RNA,extracted by use of the method,was used for electrophoresis,it clearly showed three RNA bands of 28S,18S,and 5S.The brightness of 28S RNA was twice as light as that of 18S RNA in the electrophoresis map.Purity analysis of total RNA was conducted with UV spectrophotometer.The value of OD260/OD280 varied from 1.8 to 2.0,which indicated that the RNA was not contaminated by proteins or phenol.The results indicated that the total RNA was complete and not degraded;and the entire procedure can be completed within 2 hrs.Conclusions The quality of RNA isolated from fresh tissue obtained by the improved method is stable,and electrophoresis method is rapid and reliable.