• T/A-based subcloning of PCR amplified DNA fragments

    Richard Powell Department of Microbiology, National University of Ireland, Galway, Ireland Frank Gannon European Molecular Biology Labora
    2010-03-05
  • Easy Subcloning

    Subcloning should be easy and fast, and work every time. The following protocols minimize the number of manipulations required to prepare DN
    2010-03-05
  • Random subclone generation

    (adapted from Bruce A. Roe, Department of Chemistry and Biochemistry, The University of Oklahoma, Norman, Oklahoma 73019 broe@ou.edu) A. So
    2010-03-05
  • PCR产物克隆方法

    平端连接 通常情况下,PCR产物可直接与平端载体DNA进行连接,但其连接效率效低。因为TaqDNA聚合酶具有非模板依赖性末端转移酶活性,能在两6条 DNA链的3'末端加上一个多余的碱基,使合成的PCR产物成为3'突出一个碱基的DNA分子。这种DNA分子的连接效率很低。由于PCR
    2010-03-05
  • CLONING FROM LOW MELT AGAROSE

    competent cells (DH5alpha at competency of 108 cfu/µl of pUC18). The transformation procedure it self is very similar to a standard CaCl2 tr
    2010-03-05
  • SUBTRACTIVE CLONING:Past, Present, and Future

    C. G. Sagerstr¨om,1 B. I. Sun,1 and H. L. Sive2 Whitehead Institute for Biomedical Research, Nine Cambridge Center, Cambridge,Massachusetts
    2010-03-05
  • Construction of homemade 'T-vectors'

    This method is after Marchuk,D.,et al.,1991,Nucl.Acids Res.19(5),pp1154. You will need: 10 x Taq buffer (Promega) Taq Polymerase (Promega
    2010-03-05
  • Linker Ligation (with T4 ) DNA Ligase

    Linker Ligation (with T4 ) DNA Ligase In a microcentrifuge tube prepare a solution of blunt ended, dephosphorylated DNA (100-500ng) in TE b
    2010-03-05
  • Clone Genes From a Phage Library

    The overall sequence of events is: • Titer and plate out phage • Lift plaques onto filters and prepare them for screening • Make a probe
    2010-03-05
  • TA Cloning

    I. Initial mixture 5 μl dd H2O 1 μl PCR product 1 μl ligation buffer 2 μl TA vector (add second to last) 1 μl ligase (add last) Incuba
    2010-03-05