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[DNA Library] YAC library - Generation of YAC-ends by Inverse-PCR (IPCR) 日期：2005-07-18 00:00:00 点击：186 好评：0
Thursday, December 04, 2003 Description Digest of YAC-DNA Ligation IPCR with internal primers Procedure Digest of YAC-DNA: YAC-DNA 5 ul 10 X buffer 3 ul restriction enzyme 3 ul H20 21 ul Total 30 ul Incubate at 37 C, 1 hr; heat inactivate...
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[DNA Library] YAC library - Pulse Field Gel Electrophoresis 日期：2005-07-18 00:00:00 点击：187 好评：0
Thursday, December 04, 2003 Description PFGE is needed for the separation of large DNA molecules, e.g. for separation of YACs from the yeast chromosomes. Using low melting point agarose the YAC can be isolated from the gel with the Gene Cl...
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[DNA Library] YAC library - Screening a YAC - library for positive clones 日期：2005-07-18 00:00:00 点击：201 好评：0
Thursday, December 04, 2003 Description The library is a L. esculentum library of VFNT Cherry and Rio Grande-PtoR. The cosmid library was made from EcoRI partially digested total genomic DNA cloned into the BamHI site of pYAC4, which was s...
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[DNA Library] YAC library - YAC DNA isolation procedures 日期：2005-07-18 00:00:00 点击：162 好评：0
Thursday, December 04, 2003 Description The library is a L. esculentum library of VFNT Cherry and Rio Grande-PtoR. The cosmid library was made from EcoRI partially digested total genomic DNA cloned into the BamHI site of pYAC4, which was s...
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[DNA Library] Transformation Ultra-Competent E. coli (Inoue Method) 日期：2005-07-18 00:00:00 点击：201 好评：0
Wednesday, November 19, 2003 Description Inoue, H., H. Nojima, and H. Okayama. 1990. High efficiency transformation of Escherichia coli with plasmids. Gene 96:23-28. Procedure 1. Inoculate from an overnight grown in LB. 2. Grow in 250 ml S...
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[DNA Library] Rubidium Chloride method for Transformation Competent E. col 日期：2005-07-18 00:00:00 点击：159 好评：0
Wednesday, November 19, 2003 Description No Description Procedure 1. Inoculate 1 ml from overnight culture into 100 ml Psi broth (scale up or down as needed). Incubate at 37 C with aeration to A550=0.48 2. Ice 15 min. 3. Pellet cells in ap...
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[DNA Library] Library cDNA Synthesis 日期：2005-07-18 00:00:00 点击：153 好评：0
Tuesday, November 18, 2003 Description 1 cDNA Synthesis N.B: During 1 cDNA synthesis, all steps should be carried while wearing gloves and all solutions should either have been DEPC-treated or purchased specifically for use with RNA. Proce...
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[DNA Library] Generation of uni-directional nested deletions in plasmid DN 日期：2005-07-18 00:00:00 点击：128 好评：0
Tuesday, November 18, 2003 Description Good Protocol Works well Procedure 1) Digest DNA with two restriction enzymes such that one end is ExoIII susceptible (blunt or 5' overhang) and the other is ExoIII resistant (5' recessed). The ExoIII...
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[DNA Library] Construction of homemade T-vectors 日期：2005-07-18 00:00:00 点击：167 好评：0
Tuesday, November 18, 2003 Description Construction of homemade 'T-vectors' Procedure 1) Digest plasmid to be used with a blunt-end restriction enzyme (I use EcoRV and pBluescript SK-). 2) Clean digestion reaction by phenol/chloroform extr...
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[DNA Library] Bacteriophage Plaque lifts 日期：2005-07-18 00:00:00 点击：215 好评：0
Tuesday, November 18, 2003 Description Plaque transfer is typically performed using Amersham gridded, nylon, Hybond-N membranes. Procedure 1) Place Hybond-N, 82mm membrane circles onto the surface of agar plates carrying bacteriophage infe...