Extraction methods of total RNA from Stylosanthes guianensis

The modified SDS phenol extraction method, Modified CTAB method, common Trizol and Column Plant RNAout were used to extract total RNA in leaf of Stylosanthes guianensis Reyan No.2, respectively. The quality and quantity of total RNA from abovementioned methods were compared to select the better methods by UV spectrometer and gel electrophoresis. This study indicated that the value of OD260 nm/OD230 nm of RNA extracted by modified CTAB method and Column Plant RNAout method were higher than 2.0 and the value of OD260 nm/OD280 nm of RNA extracted by modified CTAB method and Column Plant RNAout were 1.85 and 1.93, respectively. Gel electrophoresis showed that RNA extracted by modified CTAB had clearer bands of 28S rRNA and18S rRNA and they did not degrade, and that RNA extracted by Column Plant RNAout had two clearer bands of 28S rRNA and18S rRNA and they did not degrade; however, RNA extracted by other two methods degraded and dispersed to some degrees. RNA extracted by modified CTAB method and Column Plant RNAout could be reversed to cDNA. The cDNA was amplified and one clear bands of actin gene fragment was observed in agarose gel. These results further demonstrated that the quality and purity of the total RNA extracted by modified CTAB and Column Plant RNAout could be applied into molecular biology experiment, and quality and purity of the total RNA by Column Plant RNAout method were better than that by modified CTAB method.