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protocols-sops 2010/11/24 21:17 protocols-sops 2010/11/24 21:20 current
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===== Plasmid DNA isolation_ alkaline lysis mini prep ===== ===== Plasmid DNA isolation_ alkaline lysis mini prep =====
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Modified from: Sambrook & Russell 2001, Kirchhof 2002 Modified from: Sambrook & Russell 2001, Kirchhof 2002
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Materials & Reagents Materials & Reagents
Alkaline lysis solution I: 50mM glucose, 25mM Tris-Cl pH 8.0, 10mM EDTA pH 8.0 Alkaline lysis solution I: 50mM glucose, 25mM Tris-Cl pH 8.0, 10mM EDTA pH 8.0
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Alkaline lysis solution II: 0.2N NaOH, 1% SDS; make fresh Alkaline lysis solution II: 0.2N NaOH, 1% SDS; make fresh
Alkaline lysis solution III: 60ml 5M potassium acetate, 11.5ml glacial acetic acid, 28.5ml ddH2O; place on ice before use Alkaline lysis solution III: 60ml 5M potassium acetate, 11.5ml glacial acetic acid, 28.5ml ddH2O; place on ice before use
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3M Sodium acetate (pH 5.2) 3M Sodium acetate (pH 5.2)
TE buffer: 10mM Tris-Cl pH 8.0, 1mM EDTA pH 8.0 TE buffer: 10mM Tris-Cl pH 8.0, 1mM EDTA pH 8.0
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Protocol Protocol
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===== DNA quantification_ using Spectrophotometer ===== ===== DNA quantification_ using Spectrophotometer =====
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Materials & Reagents Materials & Reagents

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