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[DNA Sequencing] DNA Sequencing Sequencing reaction 日期：2005-07-18 00:00:00 点击：246 好评：0
Tuesday, November 18, 2003 Description Sequencing reaction Procedure 1) Denature dsDNA by the addition of 8ul DNA (approximately 3ug) to a sterile microcentrifuge tube containing 2ul freshly made 2M NaOH vortexed briefly and incubate at ro...
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[DNA Sequencing] Measurement of DNA concentration using PanVera fluorescence 日期：2005-07-18 00:00:00 点击：196 好评：0
Tuesday, November 18, 2003 Description Measurement of DNA concentration using PanVera fluorescence assay Procedure This Dye measures DNA concentration using a fluorescent dye which binds only double stranded DNA. The fluorescence of this d...
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[DNA Sequencing] In vitro Mutagenesis with dut ung single stranded DNA 日期：2005-07-18 00:00:00 点击：139 好评：0
Tuesday, November 18, 2003 Description In vitro Mutagenesis with dut ung single stranded DNA Procedure In an 0.5 ml microfuge tube add: 0.2 microgram single stranded dut ung DNA 1.3 microliter kinased oligo from above (33 ng) 1 microliter...
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[DNA Sequencing] Replication timing using transient hemimethylation 日期：2005-07-18 00:00:00 点击：164 好评：0
Tuesday, November 18, 2003 Description Replication timing using transient hemimethylation Procedure 1. Grow ~330 ml of RM14-3a expressing dam methylase (RM14-3a Meth5) to an OD660 of about 0.3 (3.85 x 106 cells/ml) at 23oC. Arrest with 200...
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[DNA Sequencing] Replication timing by density transfer 日期：2005-07-18 00:00:00 点击：172 好评：0
Tuesday, November 18, 2003 Description Replication timing by density transfer Procedure 1. Grow cells at least 7 generations in 0.1% (w/v) [13C]-acetate or glucose 0.01% (w/v) [15N]-ammonium sulfate Amino acids/supplements (as required, ad...
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[DNA Sequencing] Replication timing by comparative hybridization 日期：2005-07-18 00:00:00 点击：194 好评：0
Tuesday, November 18, 2003 Description Replication timing by comparative hybridization Procedure 1. Grow the cells in minimal medium with 1% Na-acetate or 3% glycerol as the carbon source, aiming for about 20 ml cells per time point. (Raff...
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[DNA Sequencing] Method for assaying gliotoxin production in Aspergillus fumi 日期：2005-07-18 00:00:00 点击：184 好评：0
Thursday, November 13, 2003 Description Method for assaying gliotoxin production in Aspergillus fumigatus Procedure Culture and extraction Culture Aspergillus fumigatus on Sabouraud agar plates for 2 days at 37C and harvest the conidia usi...
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[DNA Sequencing] A rapid method for generating gene deletions in Aspergillus 日期：2005-07-18 00:00:00 点击：180 好评：0
Thursday, November 13, 2003 Description A rapid method for generating gene deletions in Aspergillus fumigatus Procedure Step 1: Introduction of target cosmid into KS272(pKOBEG) The E. coli strain KS272(pKOBEG) is transformed with the relev...
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[DNA Sequencing] Random Primed Labeling of DNA 日期：2005-07-18 00:00:00 点击：161 好评：0
Tuesday, October 21, 2003 Description Random Primed Labeling of DNA Procedure 1. Denature 30 to 50 ng of DNA by boiling for 2 min. 2. To a microcentrifuge tube add in this order: ddH2O to give a final volume of 50 ul 10 ul of OLB Denatured...
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[DNA Sequencing] Labeling Oligonucleotides Using T4 Polynucleotide Kinase 日期：2005-07-18 00:00:00 点击：150 好评：0
Tuesday, October 21, 2003 Description Labeling Oligonucleotides Using T4 Polynucleotide Kinase Procedure 1. Add the following to a microcentrifuge tube: 2 l of 10X Kinase Buffer 3 l of -[32P]-ATP (7000 Ci/mmole or 167 Ci/ul) (CAUTION! See...