摘要:reagents: DNA for labeling (concentration c > 150 ng/µl) modified nucleotides: Biotin-16-dUTP, Digoxigenin-11-dUTP, conc. 1nmol/µl (Boehringer Mannheim) dNTPs (regular nucleotides): dATP, [阅读全文:]
摘要:This is a question asked quite a bit, most recently (late September, 1995) in the newsgroup. Here are two methods, and who submitted them. UV Shadowing Electrophorese the oligo via PAGE. Place the gel[阅读全文:]
摘要: 1. Cut DNA with 5'- and 3'- overhangs, gel check, phenol-Sevag extract and NaOAc/EtOH ppt. 2. Prepare one S1 nuclease tube (on ice) for each time point: 15ml S1 Buffer + 0.25U S1/ml (5U). 3. Use 5ml [阅读全文:]
摘要:Prepare a 100 µl reaction mixture containing the DNA of interest in the appropriate 1X restriction enzyme buffer. Divide the reaction mixture between five prechilled microcentrifuge tubes such t[阅读全文:]
摘要:基因是细胞内DNA分子上具有遗传效应的特定核苷酸序列的总称,是具有遗传效应的DNA分子片段。基因控制蛋白质合成,是不同物种以及同一物种的不同个体表现出不同的性状的根本原因,即所谓"种瓜得瓜,种豆得豆","一母生九子,[阅读全文:]
摘要:Materials:10X restriction enzyme buffer (see manufacturer's recommendation)DNAsterile waterrestriction enzymephenol:chloroform (1:1) 1.Add the following to a microfuge tube:2 μl of appropriate 10X [阅读全文:]
摘要:功能克隆(funetional cloning)是利用疾病已知的遗传损伤而引起的生化功能如蛋白质基酸缺陷的信息,进行基因定位,进而克隆 该致病基因。以进行的基因克隆 大都是预先测知疾病基因的编码蛋白质,利用其mRNA反转录成cDNA,[阅读全文:]
摘要:1 国内外研究现状 拟南芥(Arabidopsis thaliana )是一种模式植物,具有基因组小(125 Mbp)、生长周期短等特点,而且基因组测序已经完成(The Arabidopsis Genomic Initiative, 2000)。同时,拟南芥属十字花科(Cruciferae),[阅读全文:]
摘要:Subcloning should be easy and fast, and work every time. The following protocols minimize the number of manipulations required to prepare DNA fragments for ligations, thereby both saving time and incr[阅读全文:]
摘要:(adapted from Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma 73019 broe@ou.edu)A. SonicationThe generation of DNA fragments by sonication is perfo[阅读全文:]