Method of Total RNA Extraction and Cloning of LFY Gene in Castanea henryi
The quality of extracted total RNA is the key to study the molecular mechanism of chinquapin later.In this study, chinquapin inflorescences were taken as the material. The study used two types of plant RNA Rapid Extraction kit, a modified trizol method and a improved CTAB method to extract the chinquapin total RNA,and used some methods to test the quality of the RNA. The total RNA obtained by improved CTAB method could significantly observe 28 S and 18 S bands and the brightness ratio was about 2 ∶ 1, DNA hardly leave, spotting holes without impurities, OD260/OD280 were close to 2, while OD260/OD2302. The RNA can be used as a template after reverse transcription to obtain LFY gene. The improved CTAB method was most appropriate, the total RNA had high yield and relatively pure, complete and could be used for subsequent reverse transcription reaction to clone.