ABSTRACT
<p><b>OBJECTIVE</b>To optimize a simple and effective method for total RNA extraction from bulblet of Fritillaria anhuiensis.</p><p><b>METHOD</b>Four methods, i. e. guanidine isothiocyanate, bentonite, modified SDS/phenol and the RNAiso plus, were used to extract total RNA from bulblet of F. anhuiensis. Then the results of the extraction were compared and analyzed by electrophoresis detection and RT-PCR verification.</p><p><b>RESULT</b>The total RNA extracted by bentonite method were clear and no dispersion, the integrity of the RNA was well, and there was no obvious contamination with DNA and other impurities, was suitable for RT-PCR test.</p><p><b>CONCLUSION</b>The bentonite method is quick, economic, and efficient for total RNA extraction from bulblet of F. anhuiensis.</p>
Subject(s)
Bentonite , Chemistry , DNA, Complementary , Electrophoresis , Fritillaria , Genetics , Guanidines , Chemistry , Isothiocyanates , Chemistry , Phenol , Chemistry , Plant Roots , Genetics , Plants, Medicinal , Genetics , RNA, Plant , Reverse Transcriptase Polymerase Chain Reaction , Sodium Dodecyl Sulfate , Chemistry , Time FactorsABSTRACT
<p><b>OBJECTIVE</b>To determine the optimal concentration and processing time of EMS mutation for suspension cells from Pinellia ternata.</p><p><b>METHOD</b>Under four EMS concentration gradients (0.1% , 0.2%, 0.4%, 0.6%) and three processing time gradients (0.5, 1.0, 2.0 h), the suspension cells of P. ternata were mutagenized.</p><p><b>RESULT AND CONCLUSION</b>The results showed that the survival rate was significantly different under the different concentrations of EMS and the different processing time. In the same processing time, the EMS concentrations were increased, but the suspension cells survival rate decreased gradually. The optimum EMS concentration for the mutagenesis was 0.4% and the best processing time was 1 hour.</p>