Reagent and liquid nitrogen using TRIzol -k high quality total RNA extracted rat pancreas.doc

 PAGE \* MERGEFORMAT 13 Reagent and liquid nitrogen using TRIzol ?k high quality total RNA extracted rat pancreas Of: Li Dongmin, Ren Wu Chao, Wang Xuan, Wang Fei Miao, Gao, Han Yan, Ning Qilan, Song Tianbao, Lu Shemin [Abstract] Objective To establish a high quality total RNA of pancreatic fast, economical and stable extraction. Methods and liquid nitrogen extraction reagent TRIzol ?k rat pancreas total RNA, total RNA content and by determination of the ratio of A260/280 and 10g / L non- denaturing agarose gel electrophoresis of total RNA quality assessment, and by RT PCR detection of insulin 1, glucagon, pancreatic @ amylase and actin expression. Results and liquid nitrogen using TRIzol ?k total RNA extracted from rat pancreas could reach 3 ~ 6g/mg pancreatic tissue, A260/280 ratio of between 1.75 to 1.89. In 10g / L non-denaturing agarose gel electrophoresis, were seen 28S and 18S bands, and in the 28S, 18S bands was seen between the cloudy shadows. rat insulin 1, glucagon, pancreatic @ amylase, actin RT PCR product electrophoresis showed clear bands were. Conclusion TRIzol ?k reagents and liquid nitrogen successfully extracted high-quality total pancreas RNA, which can be used for RT PCR study. [Keywords:] pancreas, total RNA extraction, TRIzol ?k reagents, liquid nitrogen ABSTRACT: Objective To establish a quick, economical and reproducible method for high quality RNA extraction from pancreas. Methods We utilized TRIzol ?k Reagent and liquid nitrogen to isolate total RNA from the rat pancreas. The RNA quality was determined by detection of its content and optic density (A) at 260/280nm, and electrophoresis in 1% non denatured agarose gel. Then reverse transcription polymerase chain reaction (RT PCR) was performed to detect expression of the pancreas specific genes. Results The content of the total RNA extracted from the rat pancreas reached 3-6g/mg pancreatic tissues, and A260/280 ratio was 1.75-1.89. Electrophoresis of the total RNA showed 2