Schistosome and Snail Recipes

iBge Medium pH 7.2

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

To prepare cBge medium, add 10 mL of heat inactivated FBS and 1 mL of 100X pen/strep to 89 mL iBge.

Note: iBge is very easily contaminated

Note: Do not freeze cBge Medium

Note: Bge cells are sensitive to the quality of FBS. Several lots of FBS may have to be screened. It has been known that investigators using Bge cell lines have soon lost it due to the unsuitability of their FBS.

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

From NIH NIAID Schistosomiasis Resource Center

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

From the lab of TP Yoshino

• 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