• PCR from bacterial colonies

    This procedure is used to analyze inserts in pUC-derived plasimids. 1. Suspend E. coli colonies harbouring plasmids in 50 microliters of TE
    2010-03-05
  • Phosphatase treatment of DNA fragments

    Richard Powell Department of Microbiology, National University of Ireland, Galway, Ireland Frank Gannon European Molecular Biology Laborat
    2010-03-05
  • Standard Ligation

    Prepare a ligation mix: Ligation Mix (2x) 10x ligase buffer 1.0 ml digested vector (0.1 mg/ml) 1.0 ml H2 O 6.0 ml total 8.0 ml di
    2010-03-05
  • 在质粒载体(plasmid vector)中进行平末端片段的克隆

    1、分别设立两个反应,用适当的限制性内切核酸酶消化1-10μg质粒DNA和外源DNA片段,使它们能产生平末端。 2、苯酚:氯仿抽提与乙醇沉淀法纯化出已被消化的载体和外源DNA片段。 3、分别用TE(pH 8.0)重新溶解纯化出的两种DNA沉淀,使终浓度为100 ng/ml。假
    2010-03-05
  • Protocols for ET recombination

    1、Oligo design (1)The 5' end (the homology arm) - choose 42 or more (we usually choose about 50) nts for the homology arms from the target
    2010-03-05
  • 基因克隆的实验方法

    Serial Analysis of Gene Expression (SAGE) SAGE is a powerful tool that allows the analysis of overall gene expression patterns with digital
    2010-03-05
  • DNA重组技术(DNA Recombination)

    一、DNA 的酶切与连接 (1)酶切反应:同质粒DNA 的鉴定,只不过是质粒DNA 换为载体DNA 。若大量酶切,则成比例增加。 (2)加2倍体积的预冷无水乙醇和1/10体积的3mol/l NaAc混匀,-20℃2h以上。 (3)15000rpm离心15min,弃上清。
    2010-03-05
  • 一步实现单拷贝到高拷贝的转换--CopyControl克隆技术

    单拷贝克隆产量低,高拷贝克隆稳定性差,一直让研究者在克隆 表达时难以选择。蛋白的表达就有诱导表达系统,可以实现人为地控制蛋白的表达时间和表达量——现在连质粒的拷贝数可以借助诱导的方法来人为控制了!Epicentre的专利技术——CopyControl克隆 系统能够让您共享单拷贝和
    2010-03-06
  • 重组DNA的分离、克隆与测序实验手册

    Introduction This manual is a compilation of many of the everyday methods used in the average molecular biology laboratory, with emphasis o
    2010-03-06
  • 未知侧翼序列扩增-DNA Walking步移法

    DNA WALKING REFERRENCEKi-Bum Park and Suk-Heung Oh(2007)Cloning, sequencing and expression of a novel glutamate decarboxylase gene from a ne
    2010-03-06