BioProtocols--sol8

commonly used solutions (2) ------ 1 ) 2 ) 3 )

SDS stock

10% or 20% (w/v) SDS. Also called sodium lauryl (or dodecyl) sulfate. To prepare a 20% (w/v) solution, dissolve 200 g of electrophoresis-grade SDS in 900 ml of H2O. Heat to 68 ?C and stir with a magnetic stirrer to assist dissolution. If necessary, adjust the pH to 7.2 by adding a few drops of concentrated HCl. Adjust the volume to 1 liter with H 2O. Store at room temperature. Sterilization is not necessary. Do not autoclave.

Use a mask when weighing this out.

20x SSC

0.3M Na(3) citrate

3M NaCl

SOB Medium: per liter:

Bacto-tryptone 20 g

Yeast extract 5 g

NaCl 0.584 g

KCl 0.186 g Mix components and adjust pH to 7.0 with NaOH and autoclave.

2 M Mg ++ stock:

MgCl 2-6H2O 20.33 g

MgSO 4 -7H2O 24.65 g

Distilled water to 100 ml. Autoclave or filter sterilize.

2 M Glucose

Glucose 36.04 g

Distilled water to 100 ml. Filter sterilize.

For SOB Medium + magnesium: Add 1 ml of 2 M Mg ++ stock to 99 ml SOB Medium.

For SOCMedium: Add 1 ml of 2 M Mg ++ stock and 1 ml of 2 M Glucose to 98 ml of SOB Medium.

3M Sodium Acetate - pH 5.2

To prepare a 3 M solution: Dissolve 408.3 g of sodium acetate-3H2O in 800 ml of H 2O. Adjust the pH to 5.2 with glacial acetic acid. Adjust the volume to 1 liter with H2O. Dispense into aliquots and sterilize by autoclaving.

Southern Solutions:

Depurination solution (for Southern blotting)

250mM HCl

Denaturation solution (for Southern blotting)

1.5M NaCl

0.5M NaOH

Neutralization solution (for Southern blotting)

1.5M NaCl

0.5M Tris-HCl, pH adjusted to 7.5

50x TAE

Prepare a 50x stock solution in 1 liter of H2O:

242 g of Tris base

57.1 ml of glacial acetic acid

100 ml of 0.5 M EDTA (pH 8.0)

The 1x working solution is 40 mM Tris-acetate/1 mM EDTA.

5X (or 10X) TBE

Prepare a 5x stock solution in 1 liter of H2O:

54 g of Tris base

27.5 g of boric acid

20 ml of 0.5 M EDTA (pH 8.0)

The pH of the concentrated stock buffer should be approx. 8.3.. Some investigators prefer to use more concentrated stock solutions of TBE (10x as opposed to 5x). However, 5x stock solution is more stable because the solutes do not precipitate during storage. Passing the 5x or 10x buffer stocks through a 0.22 um filter can prevent or delay formation of precipitates.

Commonly used solutions (1)--- 1 ) 2 ) 3 ) 4 ) 5 ) 6 )

Commonly used solutions (2)--- 1 ) 2 ) 3 )

Solutions for molecular cloning---1 ) 2 ) 3 ) 4 ) 5 ) 6 ) 7 ) 8 ) 9 )