Schistosome and Snail Recipes

iBge Medium pH 7.2

• Filter sterilize (0.22 um filter) and store at 4C

Hansen's Buffered Saline pH 7.5-7.7

Chernin's Balanced Salt Solution (CBSS)

• Filter sterilize (0.22 um filter) and store at 4C
• To make CBSS+, add 1 g each of glucose and trehalose

Trypsin Solution 0.05%

• Filter sterilize (0.22 um filter).
• Freeze in 10 mL aliquots.
• Warm before using.

NIH Pond Water Stock Solution

1. Prepare 5 stock solutions in plastic Nalgene bottles:

Note: use deionized water for all solutions

1. FeCl$_3$ - 6 H$_2$O — 0.25 g/L
2. CaCl$_2$ - 2 H$_2$O — 12.9 g/L
3. MgSO$_4$ - 7 H$_2$O — 10 g/L
4. 1 N NaOH - 40 g/L
5. Phosphate buffer:
1. Dissolve 34 g KH$_2$PO$_4$ in 500 mL H$_2$O
2. Add ~175 mL 1 N NaOH until pH 7.2 is reached
3. Add 1.5 g (NH$_4$)$_2$SO$_4$
4. Add H$_2$O to 1 L
2. For 20 L pond H$_2$O:
1. 10 mL FeCl$_3$ solution
2. 50 mL CaCl$_2$
3. 50 mL MgSO$_4$
4. 25 mL phosphate buffer
3. For 50 gal
1. 95.6 mL FeCl$_3$ solution
2. 473.1 mL CaCl$_2$
3. 473.1 mL MgSO$_4$
4. 236.6 mL phosphate buffer

Yoshino Pond Water Stock Solution

• Pond water stocks are prepared in small Nalgene containers.
• Shake the stock bottle well before diluting, as the salts may have settled on the bottom.
• Common working Pond Water dilutions:
• 100 mL Pond Water Stock → 30 gallons dH2O
• 33 mL Pond Water Stock → 10 gallons dH2O
• 16.5 mL Pond Water Stock → 5 gallons dH2O
• 12 mL Pond Water Stock → 10 liters dH2O
• 1.2 mL Pond Water Stock → 1 liter dH2O