Abstract:

In order to obtain an ideal total RNA extraction method of antagonistic yeast Pichia membranefaciens, Three methods, including CTAB method, Trizol method and a modified SDS method were used to extract total RNA from yeast cell and the quality and quantity were compared with ultraviolet spectrophotometer, gel electrophoresis and RT-PCR reaction. The results indicated that the modified SDS method was better than the other methods since the value of OD260/OD280 and OD260/OD230 were 1.980 and 2.054, the purities were higher, the bands of 28S, 18S and 5S rRNA on gel electrophoresis were very clear and did not degraded, the brightness of 28S bands was twice of the bands of 18S，and the special band was amplified by RT-PCR. Comparably, CTAB method was not very good as the modified SDS method because the brightness of 28S bands was similarly as the bands of 18S. Trizol method was not fit the requirement of RNA extraction, the total RNA degraded and dispersed, DNA pollution was also found on gel electrophoresis. Therefore, the modified SDS method was appropriate for RNA extraction of antagonistic yeast and the quantity was highest.