Solutions

Puck's Saline G
vol (for 1 liter) of stock mg/liter
CaCl2.2H2O (0.11 mM) 1.1 ml 100 mM 16
MgSO4.7H2O (0.62 mM) 6.2 ml 100 mM 154
KCl (5.36 mM) 1.8 ml 3 M 400
KH2PO (1.1 mM) 11 ml 100 mM 150
NaCl (137 mM) 8000
Na2HPO4.7H2O (1.08 mM) 10.8 ml 100 mM 290
glucose 1100
phenol red 5 ml 0.1% 5

Dissolve above compounds in ~900 ml, bring to pH 7.0. Add distilled H2O and bring volume to 1 liter. Filter through sterile Millipore filter into sterile medium bottles. Store at 4oC.

Hanks' Balanced-Salt Solution
vol (for 1 liter) of stock mg/liter
NaCl (137 mM) 8000
KCl (5.36 mM) 1.8 ml 3 M 400
CaCl2.2H2O (1.26 mM) 12.6 ml 100 mM 185
MgCl2.6H2O (0.5 mM) 5 ml 100 mM 100
MgSO4.7H2O (0.4 mM) 4 ml 100 mM 100
Na2HPO4.7H2O (0.34 mM) 3.4 ml 100 mM 90
KH2PO4 (0.44 mM) 4.4 ml 100 mM 60
NaHCO3 (4.2 mM) 350
Glucose 1000
Phenol Red 10 ml 0.1% 10

Dissolve above compounds in ~900 ml, bring to pH 7.0. Add distilled H2O and bring volume to 1 liter. Filter through sterile Millipore filter into sterile medium bottles. Store at 4oC.

Phosphate Buffered Saline (Dulbecco)--Solution A
vol (for 1 liter) of stock mg/liter
NaCl (137 mM) 8000
KCl (2.68 mM) 0.9 ml 3 M 200
Na2HPO4.7H2O (8 mM) 2160
KH2PO4 (1.47 mM) 14.7 ml 100 mM 200
phenol red (optional) 10 ml 0.1% 10

Dissolve above compounds in ~900 ml, bring to pH 7.0. Add distilled H2O and bring volume to 1 liter. Autoclave (10 lb/in2 for 15 min) or filter through sterile Millipore filter into sterile medium bottles. Store at 4oC. Can be used by itself.

Most of the time, Solution A can be used by itself. To add Calcium and Magnesium, do the following.

Prepare Solution A, bring the final volume to 800 ml instead of 1000 ml.

Solution B
vol (for 100 ml) of stock mg/100 ml
CaCl2 (9 mM) 9 ml 100 mM 133 (2H2O)

Solution C
vol (for 100 ml) of stock mg/100 ml
MgCl2 (5 mM) 5 ml 100 mM

Prepare and sterilize each solution separately. Store at room temperature. To use: Add 8 parts of A to 1 part B and 1 part C.

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