Created
Jun 7, 2022 8:38 PM
Status
Complete
Category
SchistosomesSnailsRecipes
Schistosome and Snail Recipes
iBge Medium pH 7.2
Reagent | Amount (per 100 mL) |
---|---|
Schneider's Drosophila Medium | 22 mL |
Lactalbumin hydrolysate | 450 mg |
Galactose | 130 mg |
Sterile DI water | 68 mL |
Gentamicin 50 mg/ml stock (Optional) | 0.2 mL |
- 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
Reagent | Amount (per 1 L) |
---|---|
NaCl | 2.8 g |
Sodium gluconate | 1.5 g |
KCl | 0.15 g |
Na2HPO4 | 0.07 g |
HCO3 | 0.05 g |
Glucose | 0.5 g |
Trehalose | 0.5 g |
Galactose | 0.5 g |
Sterile DI water | 1 L |
Chernin's Balanced Salt Solution (CBSS)
Reagent | Amount (per 1 L) |
---|---|
NaCl | 2.8 g |
KCl | 0.15 g |
Na2HPO4 | 0.07 g |
MgSO4 heptahydrate | 0.45 g |
CaCl dihydrate | 0.53 g |
NaHCO3 | 0.05 g |
Sterile DI water | 1 L |
- 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%
Reagent | Amount |
---|---|
Trypsin | 50 mg |
Hansen's buffered saline | 100 mL |
- 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
- Prepare 5 stock solutions in plastic Nalgene bottles:
- FeCl - 6 HO — 0.25 g/L
- CaCl - 2 HO — 12.9 g/L
- MgSO - 7 HO — 10 g/L
- 1 N NaOH - 40 g/L
- Phosphate buffer:
- Dissolve 34 g KHPO in 500 mL HO
- Add ~175 mL 1 N NaOH until pH 7.2 is reached
- Add 1.5 g (NH)SO
- Add HO to 1 L
- For 20 L pond HO:
- 10 mL FeCl solution
- 50 mL CaCl
- 50 mL MgSO
- 25 mL phosphate buffer
- For 50 gal
- 95.6 mL FeCl solution
- 473.1 mL CaCl
- 473.1 mL MgSO
- 236.6 mL phosphate buffer
Note: use deionized water for all solutions
Yoshino Pond Water Stock Solution
From the lab of TP Yoshino
Reagent | Amount (per 1 L) |
---|---|
CaCO3 | 12.5 g |
MgCO3 | 1.25 g |
NaCl | 1.25 g |
KCl | 0.25 g |
Sterile DI water | 1 L |
- 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