Abstract:

To establish an efficient method for high quality total RNA extraction from leaves and fruits of G. jasminoides, total RNAs were extracted with a high salt, low pH method and an EASYspin Plant RNA Mini Kit. The quality of the RNAs was estimated by UV spectrophotometric analysis, detected by 1% non-denaturing agarose gel electrophoresis. And the products reverse-transcribed from the total RNAs were also checked by 0.7% non-denaturing agarose gel electrophoresis. Both the RNA extracted from fruits with the high salt, low pH method and the RNA extracted from leaves with the kit had given a satisfactory yield. When the above two total RNA were run on a 1% denaturing agrose gel, sharp and clear 28S and 18S rRNA bands were observed, and the robust of 28S rRNA band was twice as intense as the 18S band, respectively. The ratio A260/A280 for the two total RNAs were from 1.9 to 2.0, and cDNA products reverse transcribed from the two intact RNAs had exhibited expected sizes from 250 bp to 5000 bp. Using these cDNA above as templates, fragment amplification of 60S ribosomal protein L18A gene was carried out successfully. Therefore, high quality total RNAs of G. jasminoides could be extracted from fruits with a high salt, low pH method, and extracted from leaves with an EASYspin Plant RNA Mini Kit.