RNA Extraction and RT-PCR detection of Dendrobium candidum / 中草药

ABSTRACT

Objective The high quality RNA from the plantlet of Dendrobium candidum was isolated efficiently.It laid a foundation to study the mechanism of molecular expression on its growth and metabolism under environmental stress.Methods The groups stimulated by sound wave and the control groups were established.Total RNA was prepared by a modified phenol-chloroform method.The content was determined by a spectrophotometer.The integrity of RNA was detected by electrophoresis with 1.0% denatured agarose gel.The total RNA obtained was amplified by RT-PCR.The differential cDNA bands were displayed on 6% denatured polyacrylamide gel by electrophoresis and silver staining.Results The ratios of total RNA A_(260)/A_(230) and A_(260)/A_(280) were 3.16 and 1.96,respectively.The yield was 0.27 ?g/mg fresh weight.The bands were clear on agarose gel and the integrity of RNA was good.The differential cDNA bands were screened with molecular weight of 240—600 bp.Conclusion The method of modified phenol-chloroform can be used to isolate RNA from D.candidium with high quality and high yield.