A Method for Rapid Isolation of High-Quality Total RNA from Wheat Seeds

Isolation of high-quality RNA is a prerequisite to study the development of wheat seeds at the gene expression level.Rapid isolation of high-quality total RNA from wheat seeds is difficult with the extraction methods reported previously.In the present study,cold phenolic method and Trizol single-step method were combined for RNA extraction,and high-quality total RNA was obtained in about five hours.The quality of total RNA was analyzed with agarose gel electrophoresis and UV spectrophotometer.Clear bands of 28S rRNA and 18S rRNA were shown in the RNA electrophoresis,and the value of OD260/OD280 was 1.90 to 2.00.Using the total RNA isolated by this method for reverse transcription,high quality of cDNA could be obtained,and clear bands were detected in subsequent cDNA-AFLP analysis.These results demonstrated that the purity and integrality of total RNA isolated with the new method were significantly satisfactory for the demands of molecular biological experiments.