Effective method for isolation of total RNA from Fagopyrum cymosum callus / 中国中药杂志

ABSTRACT

<p><b>OBJECTIVE</b>To establish a method for isolation of the total RNA from Fagopyrum cymosum callus.</p><p><b>METHOD</b>The improved method combining that of CTAB extraction with the LiCl precipitation was used to isolate the total RNA from the four F. cymosum callus. The quality of the RNA was detected by UV spectrophotometric analysis, 0.8% non-denaturing agarose gel electrophoresis and RNA reverse transcription.</p><p><b>RESULT</b>The bands of 28S and 18S could be seen clearly by agarose gel electrophoresis, and the value of A260/A280 was between 1.9 and 2.0. The cDNA which was reverse-transcribed by the total RNA showed a wide length rage of 500 bp-5 kb.</p><p><b>CONCLUSION</b>The RNA extracted by this method meets the requirement of reverse transcription-polymerase chain reaction (RT-PCR), construction of cDNA libraries, et al. This improved method can be used to isolate the total RNA from F. cymosum callus with the advantage of simpleness, efficiency and low cost.</p>