IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-7-25: Difference between revisions
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* goal: to determine the minimum concentration of streptavidin that can be visualized with silver stain, and to note the times at which different concentrations become visible | * goal: to determine the minimum concentration of streptavidin that can be visualized with silver stain, and to note the times at which different concentrations become visible | ||
* methods: 12% PA gel with lanes listed below, run for 20 min. at 130 V, then stained with [http://bio-rad.com/B2B/BioRad/product/br_category.jsp?BV_SessionID=@@@@0044032730.1153852982@@@@&BV_EngineID=cccdaddiglfhhimcfngcfkmdhkkdfll.0&categoryPath=%2fCatalogs%2fLife+Science+Education%2fEquipment+and+Supplies%2fProteins+and+Proteomic+Studies%3a+Equipment+and+Reagents%2fProtein+Stains%2fSilver+Stain+Plus+Kit&catLevel=6&divName=Corporate&loggedIn=false&lang=English&country=HQ&catOID=-19568&isPA=false&serviceLevel=Lit+Request Bio-Rad Silver Stain Plus Kit] and [http://bio-rad.com/LifeScience/pdf/Bulletin_9056.pdf protocol]. | * methods: 12% PA gel with lanes listed below, run for 20 min. at 130 V, then stained with [http://bio-rad.com/B2B/BioRad/product/br_category.jsp?BV_SessionID=@@@@0044032730.1153852982@@@@&BV_EngineID=cccdaddiglfhhimcfngcfkmdhkkdfll.0&categoryPath=%2fCatalogs%2fLife+Science+Education%2fEquipment+and+Supplies%2fProteins+and+Proteomic+Studies%3a+Equipment+and+Reagents%2fProtein+Stains%2fSilver+Stain+Plus+Kit&catLevel=6&divName=Corporate&loggedIn=false&lang=English&country=HQ&catOID=-19568&isPA=false&serviceLevel=Lit+Request Bio-Rad Silver Stain Plus Kit] and [http://bio-rad.com/LifeScience/pdf/Bulletin_9056.pdf protocol]. | ||
** 35 min. in fixative solution | |||
** 2 x 10 min. wash in water | |||
** XX min. in staining solution | |||
** 15 min. in stopping solution | |||
* lanes: | * lanes: | ||
{| {{table}} | {| {{table}} | ||
Revision as of 14:26, 25 July 2006
Silver staining experiments
- goal: to determine the minimum concentration of streptavidin that can be visualized with silver stain, and to note the times at which different concentrations become visible
- methods: 12% PA gel with lanes listed below, run for 20 min. at 130 V, then stained with Bio-Rad Silver Stain Plus Kit and protocol.
- 35 min. in fixative solution
- 2 x 10 min. wash in water
- XX min. in staining solution
- 15 min. in stopping solution
- lanes:
| lane | streptavidin | water (μL) | 10x Tris-gly loading dye (μL) | |
| (μL) | (fmols) | |||
| 1 | 10 μL 4 μM | 40,000 | 0 | 2 |
| 2 | 10 μL 2 μM | 20,000 | 0 | 2 |
| 3 | 5 μL 2 μM | 10,000 | 5 | 2 |
| 4 | 5 μL 1 μM | 5,000 | 5 | 2 |
| 5 | 5 μL 0.5 μM | 2,500 | 5 | 2 |
| 6 | 2 μL 0.5 μM | 1,000 | 8 | 2 |
| 7 | 1 μL 0.5 μM | 500 | 9 | 2 |
| 8 | 2.5 μL 0.1 μM | 250 | 7.5 | 2 |
| 9 | 1 μL 0.1 μM | 100 | 9 | 2 |
| 10 | 1 μL 0.05 μM | 50 | 9 | 2 |
