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. Author manuscript; available in PMC: 2020 Aug 21.
Published in final edited form as: J Vis Exp. 2020 May 23;(159):10.3791/61243. doi: 10.3791/61243

Table 1:

Recipes

M9 buffer (1 L)
KH2PO4 3 g
Na2HPO4 6 g
NaCl 5 g
1 M MgSO4 1 mL
ddH2O up to 1 L
2x Lysis buffer (5 mL)
HEPES (pH 7.4) 200 μL
2 M KCl 250 μL
10% TritonX 100 μL
1 M MgCl2 20 μL
100% glycerol 1 mL
ddH2O up to 5 mL
Add fresh:
1 M DTT 20 μL
EDTA-free protease inhibitor 1 tablet
phosphatase inhibitor cocktail 2 100 μL
phosphatase inhibitor cocktail 3 100 μL
1x Lysis buffer
Dilute 2x Lysis buffer with an equal volume of ddH20.
1x Wash buffer 10 mL)
HEPES (pH 7.4) 300 μL
2 M KCl 500 μL
10% TritonX 100 μL
1 M MgCl2 20 μL
100% glycerol 1 mL
ddH2O up to 10 mL
1 M DTT 20 μL (add fresh)