Northern Solutions
- Dissolve 1.16 g of agarose in 62mL of milliQ H2O
- Cool agarose to 55-60 °C in a pre-warmed water bath
- Add 8.5 mL of 10x MOPs
- Add 14.45 mL of 37% formaldehyde in the hood
- Pour gel immediately and allow to cool of at least 30 min in the hood
20x SSC
| Final Concentration
|
For 1 Liter
|
For 4 Liters
|
| 3M NaCl
|
175.3 g
|
701.2 g
|
| 0.3M NaCitrate
|
88.2 g
|
352.8 g
|
| milliQ H2O
|
to 1 L
|
to 4 L
|
10x MOPs
| Final Concentration
|
For 500 mL
|
For 1 Liter
|
| 0.4 M Morpholinopropanesulfonic (MOPS) acid (free acid)
|
41.9 g MOPS
|
83.8 g MOPS
|
| 0.1 M Na-acetate-3 x H2O
|
6.8 g
|
13.6 g
|
| 10 mM EDTA
|
10 mL 0.5 M EDTA
|
20 mL 0.5 M EDTA
|
| Adjust pH to 7.2 with NaOH
|
| Store in the Dark at 4 °C
|
100x Denhardt's
| Final Concentration
|
For 100 mL
|
For 500 mL
|
| Ficoll 400
|
2 g
|
10 g
|
| Polyvinylpyrrolidone MW 360000
|
2 g
|
10 g
|
| BSA fraction V
|
2 g
|
10 g
|
| milliQ H2O
|
| Store at -20 °C
|
Hybridization Buffers
Oligo (Pre)Hybridization Buffer
| Final Concentration
|
For 100 mL
|
For 200 mL
|
| 3x SSC
|
15 mL of 20x SSC
|
30 mL of 20x SSC
|
| 0.5% SDS
|
5 mL of 10% SDS
|
10 mL of 10% SDS
|
| 1x Denhardt's
|
2 mL of 50x Denhardt's
|
4 mL of 50x Denhardt's
|
| 100 μg/mL ssDNA
|
1 mL 10 mg/mL ssDNA
|
2 mL 10 mg/mL ssDNA
|
| milliQ H2
|
to 100 mL
|
to 200 mL
|
Hyb Buffer
| Final Concentration
|
For 50 mL
|
For 100 mL
|
| PIPES
|
2.5 mL of 1M PIPES pH 6.5
|
5 mL of 1M PIPES pH 6.5
|
| Sodium Phosphate Buffer
|
2.5 mL of 1M NaPO4 Buffer pH 7.0
|
5 mL of 1M NaPO4 Buffer pH 7.0
|
| Sodium Chloride
|
1 mL of 5M NaCl
|
2 mL of 1 mL of 5M NaCl
|
| SDS
|
12.5 mL of 20% SDS
|
25 mL of 20% SDS
|
| EDTA
|
250 μL of 0.5M EDTA pH 8.0
|
500 μL of 0.5M EDTA pH 8.0
|
| ssDNA
|
300 μL 10 mg/mL ssDNA
|
600 μL 10 mg/mL ssDNA
|
| milliQ H2
|
to 50 mL
|
to 100 mL
|
Oligo Wash Buffer
| Final Concentration
|
For 500 mL
|
For 1 L
|
| 2x SSC
|
50 mL of 20x SSC
|
100 mL of 20x SSC
|
| 0.1% SDS
|
5 mL of 10% SDS
|
10 mL of 10% SDS
|
| milliQ H2
|
to 500 mL
|
to 1 L
|
Loading Buffers
MOPs Loading Buffer
| Final Concentration
|
For 1 mL
|
For 10 mL
|
| 1x MOPs
|
100 μL
|
1 mL
|
| 18.5% Formaldehyde
|
185 μL
|
1.85 mL
|
| 50% Formamide
|
500 μL
|
5 mL
|
| 4% Ficoll 400
|
40 mg
|
400 mg
|
| milliQ H2
|
215 μL
|
2.15 mL
|
| pinch of Bromophenol Blue
|
| Store in the dark at -20 °C
|
| Final Concentration
|
For 1 mL
|
For 10 mL
|
| >99% Formamide
|
0.96 mL of deionized Formamide
|
9.6 mL of deionized Formamide
|
| 20 mM EDTA
|
4 μL of 0.5M EDTA
|
400 μL of 0.5M EDTA
|
| Pinch of Xylene Cyanol and Bromophenol Blue
|
| Store in the dark at -20 °C
|
