Cell Bio Lab calculations 1 Your lab manager needs you to ma

Cell Bio Lab calculations:

1) Your lab manager needs you to make a 0.5 mg/mL Bovine Serum Albumin (BSA) in water for a Bradford Assay. At your bench, you have a 2 mg/mL BSA stock. Make 800 L of the desired solution.

2) You collected 900 L of 2 mg/mL BSA in water after a diafiltration step. Now you need to use this solution to make 200 L of protein B at 1.25 mg/mL in 1X buffer. Using the 2 mg/mL BSA stock and the 10X buffer, make the desired solution.

3) Prepare 25 mL of destain solution: 20% methanol, 10% acetic acid; 70% water.

4) To perform a cell count with the hemacytometer, you need at least 20 L of solution, typically 10 L of cell suspension and 10 L of Trypan Blue. This is referred to as a 1:2 dilution. However, when the cell density is too high to count efficiently, the solution can be further diluted with PBS. Adding to the 20 L of cell count solution at your bench, make a 1:3 and 1:10 dilution with PBS.  

Solution

1) Your lab manager needs you to make a 0.5 mg/mL Bovine Serum Albumin (BSA) in water for a Bradford Assay. At your bench, you have a 2 mg/mL BSA stock. Make 800 L of the desired solution.

Answer:

Stock = 2 mg/mL

Required = 0.5 mg/mL

Volume = 800uL =0.8 mL

C1V1= C2V2

(stock)   (required)

V1=C2V2/C1

V1 = (0.5 X 0.8 mL )/ 2

=0.2 mL

0.2 mL of stock BSA + 0.6 mL of H2O gives 800 uL of 0.5mg/mL BSA.

3) Prepare 25 mL of destain solution: 20% methanol, 10% acetic acid; 70% water.

Answer:

20% methanol means 20 mL of methanol in total of 100 mL of H20. When we make 25 mL we need ¼ th of what required for 100 mL

20% methanol, = 20/4 = 5 mL

10% acetic acid; = 10/4 =2.5 mL

70% water.=70/4 = 17.5 mL

Total = 25 mL