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Alkaline Lysis - Solution 1, 2, 3

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Biological and Medical Sciences > Classification by Description > Reagents, Kits and Assays > Media and Solutions > Solutions

Branch: Protocol-Wiki
Type: Solution Recipe
Intention: For Publication
Read/Write Permissions: Open Access
Authors :
16.06.2010 - John Stewart   

Used in Midi- or Maxi- preparations of plasmid DNA.

[edit] Solution1

25mM Tris.HCl pH8, 10mM EDTA pH8, 15% sucrose/glucose

To make 1 liter, add and mix:

  1. 600 mls of distilled water;
  2. 25ul 1M Tris-Hcl pH8;
  3. 20 ul 0.5M EDTA pH8;
  4. 150g Sucrose; and
  5. Distilled water to 1l.

[edit] Solution 2

0.5M NaOH; 1% SDS
To make 1 liter, add while mixing gently:

  1. 930mls of distilled water;
  2. 20mls 10N NaOH (mix); and
  3. 50mls of 20% SDS.

SDS can precipitate out of solution, and should not then be used.

[edit] Solution 3

3M Potassium Acetate pH5

To make 500 mls, add while mixing:

  1. 300 mls of distilled water;
  2. 294g KAc;
  3. About 140mls glacial acetic acid (pH to 5); and
  4. Distilled water to 500mls.

This solution works most effectively at stopping lysis when stored in the refridgerator and added cold.

None of these solutions require autoclaving for the purpose of plasmid preparation, and can be kept for up to 6 months at room temperature.

  • This page was last modified 20:33, 16 June 2010.
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