Construction of BAC Libraries：SOLUTIONS FOR BAC LIBRARY CONSTRUCTION

 

 

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%