
Overview:
UBE2C or ubiquitin-conjugating enzyme E2C is a member of the E2 ubiquitin-conjugating enzyme family which is required for the destruction of mitotic cyclins and for cell cycle progression (1). UBE2C acted as a dominant-negative inhibitor which blocks the destruction of mitotic cyclins A and B and the onset of anaphase in frog embryos and mammalian cells (2). High UBE2C mRNA expression is associated with poor disease-free survival and overall survival. High tumor grade, as well as high Ki67 protein expression, was more frequent in the high-expression group of UBE2C.
Gene Aliases:
dJ447F3.2; UBCH10
Genbank Number:
NM_007019
References:
1. Rape, M. et.al: Autonomous regulation of the anaphase-promoting complex couples mitosis to S-phase entry. Nature 432: 588-595, 2004. 2. Townsley, F. M. et.al: Dominant-negative cyclin-selective ubiquitin carrier protein E2-C/UbcH10 blocks cells in metaphase. Proc. Nat. Acad. Sci. 94: 2362-2367, 1997.
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植物材料 用最精确的方法,称取不超过100mg的植物材料,置于处理过的研钵,加入液氮进行研磨 将研磨得到的粉末,快速转移至无RNase,并经过液氮冷却的2mL离心管(自备),
但是,这样做的前提的,RNA提得要好,浓度要高,这样免得RNA降解到不能用了。因为在有二价阳离子的(DNAseI buffer含有)情况下,RNA在60度以上时必然发生非酶促降解。
想搞清一个问题,如何区别是因为small RNA 引起的调控呢?还是tRNA引起的调控,有没有有效的分离这两种RNA的办法呢?
样品处理:
a. 植物组织:取新鲜或-70℃冻存100mg组织在液氮中研磨,把粉末加入到1ml裂解液中混匀。
b. 动物组织:取新鲜或-70℃冻存100mg组织加1ml裂解液,用组织研磨杵或匀浆器匀浆处理。
c. 贴壁细胞:直接在培养板中加入裂解液裂解细胞,每106细胞加1ml 裂解液。用取样器吹打混匀。

