Establishment of the total RNA extraction system for lily bulbs with abundant polysaccharides

Abstract

Isolating high quality RNA from lily bulbs is made difficult by the presence of plentiful polysaccharides. To solve this problem, an efficient and steady RNA extraction technique from different parts of scales in three hybrids lily bulbs has been developed through modifications of three common RNA extraction methods, cetyltrimethylammonium bromide (CTAB), sodium dodecyl sulfate (SDS) and Trizol method. NaCl concentration of both CTAB and SDS protocols was increased in order to get rid of the polysaccharides in scales. The resulting RNA is of high-quality, as judged by agarose gel electrophoresis and UV-spectroscopic analysis. Furthermore, the middle scales obtain the highest RNA yield followed by the exterior and the interior scales. However, Trizol method was also improved to remove polysaccharides but failed. The results of reverse-transcription-polymerase chain reaction (RTPCR) indicated that only the improved CTAB method is suitable for subsequent purposes, such as reverse transcription PCR and cDNA library construction.

Key words: Lilium bulbs, polysaccharides-rich tissues, RNA isolation, improved cetyltrimethylammonium bromide (CTAB) method.