Construction of BAC Libraries：SOLUTIONS FOR BAC LIBRARY CONSTRUCTION
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SOLUTIONS FOR BAC LIBRARY CONSTRUCTION
10X Homogenization Buffer (HB) stock: (1 liter)
| Ingredient | Amount | Final Concentration |
|---|---|---|
| Trisma base | 12.1 g | 0.1 M |
| KCl | 59.7 g | 0.8 M |
| EDTA | 37.2 g | 0.1M |
| Spermidine-HCL | 2.55 g | 10 mM |
| Spermine-HCL | 3.48 g | 10 mM |
-
Adjustment: pH to 9.4-9.5 with NaOH. Store at 4 °C.
1X Homogenization Buffer (HB): (1 liter)
| Ingredient | Amount | Final Concentration |
|---|---|---|
| 10X HB stock | 100 ml | 1X |
| Sucrose | 171.15 g | 0.5M |
-
Store at 4 °C.
Adjustment: Before use add beta-mercaptoethanol to 0.15%.
20% Triton-X100 in 1 X HB: (100 ml)
| Ingredient | Amount | Final Concentration |
|---|---|---|
| 10X HB stock | 10 ml | 1X |
| Sucrose | 17.12 g | 0.5M |
| Triton X-100 | 20 ml | 20% |
-
Store at 4 °C.
Wash Buffer (1X Homogenization Buffer (HB) plus 0.5 % Triton X-100): (1 liter)
| Ingredient | Amount | Final Concentration |
|---|---|---|
| 10X HB stock | 100 ml | 1X |
| Sucrose | 171.15 g | 0.5M |
| Triton X-100 (20% stock in 1 X HB) | 25 ml | 0.5% |
