Oneill Lab:WMISH Chemicals: Difference between revisions
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Revision as of 09:16, 2 October 2007
Michael J. O'Neill Lab University of Connecticut Department of Molecular and Cell Biology
WMISH Solutions
Post-Fix
This solution must be made fresh. Glutaraldehyde is a potent fixative, and should only be used in a chemical fume hood. Do not dispose of excess fix or glutaraldehyde solution in sink.
Ingredients | 5ml | 10ml | 15ml | 20ml |
---|---|---|---|---|
37% Formaldehyde | 915.5 μL | 1.831 mL | 2.7465 ml | 3.662 ml |
25% Glutaraldehyde | 20 μL | 40 μL | 60 μL | 80 μL |
PBS + 0.1% Tween20 | 4.0645 mL | 8.121 mL | 12.1935 mL | 16.258 mL |
1M NaOH | 4.5 μL | 9 μL | 13.5 μL | 18 μL |
Hybridization Buffer
Hyb buffer should be made fresh for each hybridization. Excess can be stored at -20°C for use in post-hyb washes. Always preheat to 70°C before use.
Ingredients | [Final] | 50mL |
---|---|---|
Formamide | 50% | 25mL |
DEPC 20x SSC | 1.3x | 6.5 mL |
DEPC 0.5M EDTA | 5mM | 0.5 mL |
10mg/ml tRNA | 50 μg/mL | 250 μL |
Tween 20 | 0.2% | 101 μL |
10% CHAPS | 0.5% | 2.5 mL |
50 mg/ml Heparin | 100 μg/mL | 100 μL |
DEPC Water | 18.3 mL |
TBST
After all materials combined, solution should be filter sterilized.
Ingredients | 500 mL |
---|---|
5M NaCl | 13.69 mL |
1M KCl | 1.34 mL |
1M Tris pH 7.5 | 12.5 mL |
Tween 20 | 505 mL |
DEPC Water | to 500 mL |