Miniprepnoflowchart - Plasmid Minipreps Mini-Prep A rapid small scale method of obtaining or retrieving plasmid DNA(plasmid DNA foreign inserted

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Unformatted text preview: Plasmid Minipreps Mini-Prep: A rapid, small scale method of obtaining or retrieving plasmid DNA (plasmid DNA + foreign/ inserted DNA) from bacterial cells. 2 General Variations: 1. Boiling Method 2. Alkaline Lysis Method (method we are using) Overall Goals: To lyse cells using Alkaline Lysis Method To precipitate out proteins and bacterial DNA using Sodium Acetate To precipitate out the plasmid DNA with Isopropanol To dry and freeze plasmid DNA for further study Important Solutions and Their Purpose: Solution 1: GTE (Glucose Tris EDTA):- Tris is a buffer that works at physiological pH (pH = 7.4).- EDTA binds divalent cations in the lipid bilayer, thus weakening the cell membrane and also protecting DNA from DNAses. Solution 2: SDS/NaOH (Sodium dodyclsulfate + Sodium Hydroxide) - Alkaline Lysis Solution :- This alkaline mixture lyses the bacterial cells. The detergent SDS dissolves the lipid component of the cell...
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This note was uploaded on 05/03/2008 for the course BIO 2322 taught by Professor Spotswood during the Spring '08 term at The University of Texas at San Antonio- San Antonio.

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Miniprepnoflowchart - Plasmid Minipreps Mini-Prep A rapid small scale method of obtaining or retrieving plasmid DNA(plasmid DNA foreign inserted

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