摘要: Based on: Wan, C.-Y. and Wilkins, T.A. 1994. Anal. Bioch. 223:7-12. 1.Collect young expanding leaves (or other tissue) and freeze in liquid N2 and store at -80℃. 2.Pulverize tissue to a fine powder i[阅读全文:]
摘要:1) Harvest 1x105 to 108 cells. Wash 1x w/PBS and freeze in liquid N2 and store @ -70℃ 2) Resuspend each pellet in 1.5ml lysis buffer (300μl 5x Lysis, 75μl 200mM VR, 1.125ml H2 O3) Incubate on ic[阅读全文:]
摘要:This is the preferred method for yeast RNA preparation use Gloves and RNAse free solutions throughout.1. Use a YPD overnight culture to innoculate fresh YPD media and grow cells at 30 degrees overnigh[阅读全文:]
摘要:实验目的 从黄化小麦苗中提取总RNA ,可以为cDNA 文库的构建做好准备。我们重点是要保证RNA 的完整性、产率、防止修饰和纯度,尤其是完整性、防止修饰和纯度。RNA 是单链,2’OH是它的化学性质远比DNA 活泼。所以,提取RN[阅读全文:]
摘要: RNA Isolation - Volumes and weights are for 10 ml cultures (1-2 x 107 cells/ml).1.Spin down cells, decant, and resuspend in 0.2 ml extraction buffer with SDS and transfer to a 1.5 ml eppendorf tube. [阅读全文:]
摘要:READ THROUGH ALL CAUTIONS BEFORE TRYING THIS EXPERIMENTMaterials•Rat liver (fasted rat)•Liquid nitrogen•p-Amino-salicic acid•Phenol mixture•Homogenizer or blender6•Refrig[阅读全文:]
摘要:T℃OS-M6 cells grow in 6-well plates, 2ml media total T℃V1PD cells grow in 100mm plates.1: Aspirate off the media from the cells and was the cell monolayers with 2x5ml (To 100mm dish) or 2x1ml (to 6-we[阅读全文:]
摘要: 第一节 概 述 从真核生物的组织或细胞中提取mRNA ,通过酶促反应逆转录合成cDNA的第一链和第二链,将双链cDNA和载体连接,然后转化扩增, 即可获得cDNA文库,构建的cDNA文库可用于真核生物基因的结构、表达和调控的分析;[阅读全文:]
摘要:真核细胞的mRNA分子最显著的结构特征是具有5’端帽子结构(m7 G)和3’端的Poly(A)尾巴。绝大多数哺乳类动物细胞mRNA的3’端存在20-30个腺苷酸组成的Poly(A)尾,通常用Poly(A+ )表示。这种结构为真核mRNA[阅读全文:]
摘要:Add 0.1 ml DEPC to 100 ml of the solution to be treated and shake vigorously to bring the DEPC into solution.Let the solution incubate for 12 hours at 37℃.Autoclave for 15 minutes to remove any trace [阅读全文:]