用TRIzol试剂一步法提取甜菜花蕾中的总RNA  

A method for extraction of total RNA from the buds of sugar beet using TRIzol reagent

作　　者：于冰[1] 李海英[1] 张绍军[1] 马春泉[1] 郭德栋[1] 王桂芝[1] 

机构地区：[1]黑龙江大学生命科学学院,黑龙江哈尔滨150080

出　　处：《黑龙江大学自然科学学报》2004年第1期138-140,共3页Journal of Natural Science of Heilongjiang University

基　　金：国家863计划项目(863-101-04一01-07);国家转基因科研计划项目(G99-A-014);黑龙江省自然科学基金重点项目(ZJN-03-6);黑龙江省普通高等学校骨干教师创新能力资助项目;黑龙江大学杰出青年基金资助项目

摘　　要：在花期分别对无融合生殖甜菜M14品系和进行正常有性生殖的二倍体甜菜A2Y品系的花蕾进行取样,采用TRIzol试剂一步法,分别提取了它们的总RNA,并测定了总RNA的浓度及纯度。讨论去除RNA酶的方法、具体操作的注意事项和TRIzol试剂一步法与其它方法相比的优越性。该方法为进一步进行mRNA差异显示分析和抑制消减杂交研究提供了总RNA的来源,也为今后能在甜菜中寻找和克隆与无融合生殖相关的基因提供了可靠的保证。M14, the apomictic monosomic addition line of Beta vulgaris, was first obtained in the world in 1995. In florescence, the buds of the M14 and B. vulgaris are taken to extract their total RNA using TRIzol reagent. The density and purity are determined. The methods of eliminating the RNase, the notes of the test and the merits of TRIzol reagent comparing with other methods are discussed. The system of extracting total RNA from sugar beet buds has been built which can offer a stable RNA resource for mRNA differential display and suppression subtractive hybridization. It can be further used to find and clone apomictic genes or relative genes. The system can provide a foundation for further study of apomictic sugar beet.

关 键 词：甜菜 无融合生殖 总RNA 

分 类 号：S566.3[农业科学—作物学] S188