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| Protocol submitted by VGP from Stukenberg lab protocols [http://www.xenbase.org/community/person.do?method=display&personId=1311&tabId=0]
| | ==Reporting a bug in Xenbase== |
| | If you find a bug in Xenbase, we would really like to know! |
| | However, it is very important that we can find your bug so that we can fix it. |
| | To help us find your bug, please fill out [http://xenbase.org/xenwiki/images/6/6e/Bug_report.txt this] form, and e-mail it to us! |
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| | | ==Bug Handling Process== |
| link to protocol page [http://people.virginia.edu/~djb6t/LabWeb/frogs.htm]
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| Silver Staining Protocol (Not Compatible with Mass Spec)
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| Notes:
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| Be careful not to touch gel during any step of the procedure with either a gloved or ungloved hand, finger etc…
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| Use MilliQ-H2O for all steps
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| Perform all incubations in well washed glass dishes.
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| 1. Remove gels from glass plates in which they were run:
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| 2. Incubate in 50% Methanol, 10% Glacial Acetic Acid, 40% H2O for 30 minutes (100 ml)
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| 3. Incubate in 5% Methanol, 7% Acetic Acid (Glacial), 88% H2O for 30 minutes (100ml)
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| 4. Incubate in 10% Gluteraldehyde made up in H2O from 25% stock (25 ml) for 30 minutes.
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| 5. Wash 4 x 20-30 minutes in H2O using 200-300 ml per wash.
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| 6. Makes 2 Solutions, A and B: A) 3 ml of 30% Ammonia Hydroxide, 0.25 g NaOH in 190 ml H2O. B) 1.2g Silver Nitrate (AgNO3) into 10 ml of H2O.
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| 7. Add solution B to A slowly, mix gently, and add to gel, incubate for 30 minutes.
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| 8. Make developer and stop solutions (see below).
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| 9. Perform 5 x 1 minute washes with H2O, using 200 ml’s per wash.
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| 10. Add developer solution (~100 ml) and wait and watch gel until you see bands come up. Quickly remove developer solution and add stop solution. (Developer Solution: 0.1g Citric Acid, 1ml 37% formaldehyde into 1L H2O. Stop Solution: 190 ml H2O and 10 ml Acetic Acid, Glacial)
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| 11. After 10 minutes in Stop Solution wash thoroughly
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Reporting a bug in Xenbase
If you find a bug in Xenbase, we would really like to know!
However, it is very important that we can find your bug so that we can fix it.
To help us find your bug, please fill out this form, and e-mail it to us!
Bug Handling Process