Alkaline Denaturation Solution
| Final Concentration
|
For 1 Liter
|
For 4 Liters
|
| 1.5 M NaCl
|
87.7.3 g
|
351 g
|
| 0.5 M NaOH
|
20 g
|
80 g
|
| milliQ H2O
|
to 1 L
|
to 4 L
|
Neutralization Solution
| Final Concentration
|
For 1 Liter
|
For 4 Liters
|
| 1.5 M NaCl
|
87.7 g
|
351 g
|
| 0.5 M Tris Base
|
60 g
|
240 g
|
| Adjust pH to 7.5 with HCl
|
| milliQ H2O
|
to 1 L
|
to 4 L
|
Pre-hybridization Buffer
| Final Concentration
|
For 100 mL
|
For 250 mL
|
| 6x SSC
|
30 mL of 20x SSC
|
75 mL of 20x SSC
|
| 2.5x Denhardt's
|
5 mL of 50x Denhardt's
|
12.5 mL of 50x Denhardt's
|
| 0.5% SDS
|
2.5 mL of 20% SDS
|
6.25 mL of 20% SDS
|
| milliQ H2O
|
to 100 mL
|
to 250 mL
|
| Store at RT or 4°C
|
| Warm up the solution prior to use to redissolve the SDS
|
Hybridization Buffer
| Final Concentration
|
For 100 mL
|
For 250 mL
|
| 6x SSC
|
30 mL of 20x SSC
|
75 mL of 20x SSC
|
| 1x Denhardt's
|
2 mL of 50x Denhardt's
|
3 mL of 50x Denhardt's
|
| 0.1% SDS
|
0.5 mL of 20% SDS
|
1.25 mL of 20% SDS
|
| milliQ H2O
|
to 100 mL
|
to 250 mL
|
| Store at RT or 4°C
|
| Warm up the solution prior to use to redissolve the SDS
|