Recipe

Zero-calcium HL-3

Component Amount to add (for 1 L) Final concentration
NaCl 4.09 g 70 mM
MgCl2•6H2O 4.07 g 20 mM
NaHCO3 0.84 g 10 mM
Trehalose 1.89 g 5 mM
Sucrose 39.36 g 115 mM
HEPES 1.19 g 5 mM
KCl (1 M) 5 mL 5 mM
Na3EGTA (100 mM, pH 9.2) 5 mL 0.5 mM
1. Add all of the dry components to ~750 mL H2O. Stir to dissolve.
2. Add the KCl and Na3EGTA solutions.
3. Adjust pH to 7.2. Do not use KOH, because it will alter [K+] in the saline.
4. Adjust the final volume to 1 L with H2O.
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  1. doi:10.1101/pdb.rec12365 Cold Spring Harb Protoc 2010: pdb.rec12365-
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