Mike Barnkob:Protocols/Cloning/Solutions

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Basic solutions for cloning work.

0.5M EDTA solution

  1. Stir 186.1g disodium ethylenediamine tetraacetate (EDTA) into 800 ml of distilled water.
  2. Stir the solution vigorously using a magnetic stirrer.
  3. Add NaOH solution to adjust the pH to 8.0.
  4. Dilute the solution to 1 L with distilled water.
  5. Filter the solution through a 0.5 micron filter.
  6. Sterilize in an autoclave

TBE buffer

Creates 10x TBE buffer solution

  1. Mix the following in flask:
    • 108g Tris Base (Trizma base)
    • 55g Boric acid
    • 40 ml 0.5M EDTA solution (pH 8.0)
  2. Fill up to 1 liter of dH20
  3. Store at room temperature.

Dilute 100ml into 900 dH20 for a 1x solution. Stable for 6 months at room temperature.

1% SDS solution

  1. Mix 10g of SDS into 1 liter of deionized water
  2. Store at room temperature.