Stock_Solution_Recipes.md

Herr Lab - Recipes for Stock Solutions

TBE (Tris-borate-EDTA) (10X)

108 gm Tris Base (Trizma) 55 g boric acid 40 ml 0.5 M EDTA (pH 8.0)

Add the above to 800 mL dH2O. Stir until dissolved. Adjust to 1 L with additional dH2O.

Store at room temperature.

0.5 M EDTA (pH 8.0)

Add: 186.1 g of disodium EDTA (Na2EDTA) 800 mL of dH2O

Adjust the pH to 8.0 with NaOH (approximately 50 ml of NaOH)

Bring volume to 1 L with dH2O

Stir vigorously on a magnetic stirrer*

Sterilize by autoclaving

Store at room temperature

The disodium salt of EDTA will not dissolve until the pH of the solution is adjusted to 8.0 by the addition of NaOH.

1 M Tris-HCl

Dissolve: 121. g Tris Base (TRIZMA) 700 ml dH2O

Add concentrated HCl to desired pH: pH 7.4 : approximately 70 mL pH 8.0 : approximately 42 mL pH 9.0 : approximately 8 mL

Fill up to volume 1 L with dH2O. Sterilize by autoclaving for 20 minutes on liquid setting. Store at room temperature.

Tris-EDTA 100X

(1.0M Tris-HCl, 0.1 M EDTA, pH 8.0) 157.6 g Tris-HCL 37.22 g EDTA

Bring the solution up to 1 Lt with H2O (before you must adjust the pH to 8 using NaOH)

If preparing only 250 ml: 39.4 g Tris-HCL 9.03 g EDTA Bring the solution up to 250 ml with H2O (before you must adjust the pH to 8 using NaOH)

EDTA 0.5 M

186.1 g EDTA Bring the solution up to 1 Lt with H2O (adjust pH to 8 with NaOH)

If preparing only 250 ml: 46.53 g EDTA Bring the solution up to 250 ml with H2O (adjust pH to 8 with appoximately 5g NaOH)

DTT (Dithiothreitol) 400mg/ml

0.4 g in 1 ml H2O If you will prepare the entire bottle of DTT (1 g) then you just add 2.5 ml of H2O

NaCl 5 M

292.2 g in 1 LT H2O

If preparing only 100 ml: 29.2 g in 100 ml H2O

5X Extraction Buffer – 40 ml Stock

2 ml Tris + EDTA (100X) 7.6 ml EDTA (0.5 M) 0.4 ml NaCl (5 M) 30 ml ddH2O

SDS (Sodium dodecyl sulfate) 10%

Dissolve: 10 g of SDS in 80 mL of H2O, and then add H20 to 100 mL

Extraction Solution Recipe (1 RXN)

Water 520 ul 5X Extraction Buffer 160 ul SDS 10% 80 ul Proteinase K 10 mg/mL 20 ul DTT 400 mg/mL 20 ul Total volume 800 ul

Bromophenol Blue Loading Dye (6X)

Note: dark blue runs faster on the gel

Recipe 1: For 100 mL: 60 g glycerol into a 100-mL graduated cylinder 12 mL 0.5 M EDTA, pH 8 Add 10 mg bromophenol blue

Bring up the volume to 100 mL with ddH2O

Recipe 2: 100 mg Bromophenol Blue 6 g of Ficoll (Type 400) Fill up to 40 ml with dH2O

Dissolve well. It takes a while. An option is to shake at 37 degrees overnight. Store at room temperature.

GelRed Gel: Use dark 1.5 ml tubes. Store stock solution at dark place and room temperature.

Loading Dye Mix

Mix: 10 µl GelRed 1000 µl Loading Dye (6X) Vortex and centrifuge Dilute in water Mix

Mix: 1 µl GelRed 100 µl ddH2O

Ladder: 1 µl Ladder 1 µl Loading Dye (6X) 8 H2O