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[DNA Sequencing] Taq-polymerase catalyzed cycle sequencing using fluorescent- 日期：2005-07-18 00:00:00 点击：277 好评：0
Thursday, November 20, 2003 Description Each base-specific fluorescent-labeled cycle sequencing reaction routinely included approximately 100 or 200 ng Biomek isolated single-stranded DNA for A and C or G and T reactions, respectively. Dou...
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[DNA Sequencing] Radiolabeled sequencing gel preparation, loading, and electr 日期：2005-07-18 00:00:00 点击：188 好评：0
Thursday, November 20, 2003 Description To prepare polyacrylamide gels for DNA sequencing, the appropriate amount of urea is dissolved by heating in water and electrophoresis buffer, the respective amount of deionized acrylamide-bisacrylam...
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[DNA Sequencing] Bst-catalyzed radiolabeled DNA sequencing For single-strande 日期：2005-07-18 00:00:00 点击：181 好评：0
Thursday, November 20, 2003 Description Bst DNA polymerase-catalyzed radiolabeled two-step sequencing reactions For single-stranded DNA sequencing Procedure 1. Prepare the following extension reaction in a microcentrifuge tube: 750 ngM13 t...
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[DNA Sequencing] DNA sequencing gels 日期：2005-07-18 00:00:00 点击：225 好评：0
Tuesday, November 18, 2003 Description sequencing gels Procedure 1) Clean the glass plates extensively with detergent and water, tap water, distilled water and finally ethanol. Wipe dry with a clean paper towel. 2) Siliconize the smaller o...
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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...