4. Two common buffers in biological research are PBS and TBS. What are the compositions of these buffers and what types of experiments are they commonly used for? Cite any sources used.
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- 1. Calculate the pH of a buffer solution made from 0.20 M CH3COOH and 0.50 M CH3COO- that has an acid dissociation constant for CH3COOH of 1.8 x 10-5.1. How do you prepare 25 mL of a 0.4X solution of SDS from a 20X stock?2. How do you prepare 50 mL of a 0.25X solution of Trisethylenediaminetetraacetic acid (TE) buffer from a 100X stock?1. What volumes of 50 mM CH3COOH and 30 mM NaCH3CO0 solutions are needed to prepare 2L of 20 mM acetate buffer pH 3.6? (pka for acetic acid is 4.74)
- 2. Calculate the pH of a buffer containing 0.1 M sodium acetate and 0.2 M acetic acid (pKa = 4.76).2. 3. Perform all the calculations necessary for the volume of stock solution or water, in mL, needed to make the dilute solutions listed below. a. b. C. Make: 200 mL of 30% Ethanol dilute solution Stock Volume needed (mL) 95% Ethanol Reagent water Make: 100mL of .09%NaCL Stock 1% NaCl Reagent water 30% SDS 3 M NaOH Make: 100 mL of a 3% SDS and 0.5 MNaOH dilute solution as follows: Stock Volume needed (mL) Reagent water 200 mL Total 1% Lactose 1% Dextrose Volume needed (mL) 100 mL Total Calculate the grams of chemical needed for each stock listed below. Concentration of Stock to Prepare Volume of Stock Needed 1% Sucrose 100 mL Total 10 mL 20 mL 25 mL Amount of chemical Needed (in grams) Your instructor will have you make ONE of the above solutions-ask your instructor which solution to make.can you show how to calculate the amount of two components for the required volume of a buffer with the receipes? Prepare enough 10X stock solution § Prepare 1LProportions§ 0.25M tris§ 1.92M glycine§ pH 8.3§ 1% (w/v) SDSInstructions to prepare 100mL of 10X running buffer1. Weigh 3.02g of Tris base and 14.42g of glycine into ~90mL of ddH2O in a clean beaker and stir todissolve.2. Check the pH with a calibrated pH metre once dissolved.§ The pH of the buffer should be 8.3. pH adjustment is not required if between 8.1-8.5.3. Add 5mL 20% (w/v) SDS stock solution (a liquid) to make a 1% (w/v) solution.4. Top the volume to 100mL by adding 5mL ddH2O from a graduated cylinder.
- If I want to make a 100mL buffer that has 2% SDS, 100mM tris pH 8, 10mM EDTA, and proteinase K [1ug/ml]; How much of each would I need to add to create the buffer?1.2. Using DEAE-cellulose as ion exchange resin, indicate the starting and ending pH for the narrowest experimental pH range used to separate an amino acid mixture consisting of Cys, His and Leu Starting pH: _____ Ending pH: _____3. A buffer was prepared by dissolving 15 grams if ammonium chloride in 100mL solution prepared by diluting 15mL of concentrated ammonia [28% by mass with a density of 1.00 g/mL] to make 100mL solution of dilute ammonia. a. What is the pH of the buffer solution? b. What will be the change in pH if 10mL of 0.005M HCl is added to 25mL of this buffer solution? c. What will be the change in pH if 10mL of 0.005M NaOH is added to 25mL of this buffer?
- You need a buff er at pH 7.5 for use in purifying a protein at 4°C. You have chosen Tris, pK 8.08, ΔH° = 50 kJ · mol−1. You carefully make up 0.01 M Tris buffer, pH 7.5 at 25°C, and store it in the cold to equilibrate it to the temperature of the purifi cation. When you measure the pH of the temperature-equilibrated buff er it has increased to 8.1. What is the explanation for this increase? How can you avoid this problem?You are preparing 1 Lt of TAE (Tris Acetate EDTA) gel running buffer from a 50x stock solution. How much 50 x TAE and how much water will you combine?3.) A mixture of amino acids was subjected to paper chromatography with six amino acid standards in order to determine its composition. After visualization using ninhydrin, the measured distances were as follows: A B с D E F G a.) What is the Rf value of each amino acid observed? X Distances: Solvent 10.3cm A 4.50cm B - 1.32cm C - 6.60cm D - 5.20cm E - 3.52cm F 8.70cm G - 10.26cm X1 - 9.72cm X2 - 4.3cm X3 7.1cm - b.) What are the possible components of the amino acid mixture? Answer can be up to two amino acids per component.