AAV纯化试剂配制和浓度

AAV碘克沙醇纯化法常用试剂配制浓度
5 M NaCl 146.1 g NaCl
Up to 500 mL Ultrapure water
1 M Tris HCl (pH 8.5) 12.11 g Tris base
Up to 100 mL Ultrapure water
Add 1 M HCl using a Pasteur pipette to reduce the pH to 8.5
Lysis buffer 15 mL of 5 M NaCl
25 mL of 1 M Tris HCl (pH 8.5)
Up to 500 mL Ultrapure water
10x Phosphate-buffered saline (PBS) 80 g NaCl
2 g KCl
14.4 g Na2HPO4
2.4 g KH2PO4
Up to 1 L ddwater
1 M MgCl2 20.33 g MgCl2-6H2O
Up to 100 mL Ultrapure water
1 M KCl 7.45 g KCl
Up to 100 mL Ultrapure water
PBS-MK: 5x PBS Magnesium-Potassium stock solution 250 mL of 10x PBS
2.5 mL of 1 M MgCl2
6.25 mL of 1 M KCl
Up to 500 mL Ultrapure water H2O
15% Iodixanol 12.5 mL of Optiprep density gradient medium(Sigma-Aldrich,D1556)
10 mL of 5 M NaCl
10 mL of 5x PBS-MK
17.5 mL of Ultrapure water
25% Iodixanol 20.8 mL of Optiprep density gradient medium
10 mL of 5x PBS-MK
19.2 mL of Ultrapure water
100 µL of phenol red
40% Iodixanol 33.3 mL of Optiprep density gradient medium
10 mL of 5x PBS-MK
6.7 mL of Ultrapure water
60% Iodixanol 50 mL of Optiprep density gradient medium
100 µL of phenol red

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