General Sample Prep

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Phosphate Buffered Saline (PBS) 10X 1. Dissolve the following in 800ml distilled H2O. 80g of NaCl 2.0g of KCl 14.4g of Na2HPO4 2.4g of KH2PO4 2. Adjust pH to 7.4. 3. Adjust volume to 1L with additional distilled H2O. 4. Sterilize by autoclaving TAE 50X 1. Add the following to 900ml distilled H2O 242g Tris base 57.1ml 1MGlacial Acetic Acid 18.6 g EDTA 2. Adjust volume to 1L with additional distilled H2O TBS 10x (concentrated Tris-buffered saline) For 1 L: 1. Add the following to 900 ml distilled H20 24 g Tris base (formula weight: 121.1 g) 88 g NaCl (formula weight: 58.4 g) 2. Dissolve in 900 mL distilled water 3. pH to 7.6 with 12 N HCl 4. Add distilled water to a final volume of 1 L For a 1x solution, mix 1 part of the 10x solution with 9 parts distilled water and adjust pH to 7.6 again. The final molar concentrations of the 1x solution are 20 mM Tris and 150 mM NaCl. TBS 10x alternative recipe (concentrated Tris-buffered saline) An alternative recipe for Tris buffer combines Tris base and Tris-HCl. This avoids the large volume of potentially hazardous hydrochloric acid that is needed to neutralize a solution of Tris base alone. For 1 L: 24 g Tris-HCl (formula weight: 157.6 g) 5.6 g Tris base (formula weight: 121.1 g) 88 g NaCl (formula weight: 58.4 g) 1. Dissolve in 900 mL distilled water 2. The pH of the solution should be about 7.6 at room temperature. If too basic, adjust to pH 7.6 with concentrated HCl, and if too acidic, adjust with concentrated NaOH. 3. Add distilled water to a final volume of 1 L. 4. For a 1x solution, mix 1 part 10x with 9 parts distilled water and pH to 7.6 again. 5. The final molar concentrations of the 1x solution are 20 mM Tris and 150 mM NaCl. Shape TBST (Tris-buffered saline, 0.1% Tween 20) For 1 L: 100 mL of TBS 10x 900 mL distilled water 1 mL Tween 20