IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-7-25: Difference between revisions

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* 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].
* lanes:
* lanes:
** 1: 10 {{ul}} 4 {{um}} streptavidin (40 pmol), 2 {{ul}} loading dye
{| {{table}}
** 2: 10 {{ul}} 2 {{um}} streptavidin (20 pmol), 2 {{ul}} loading dye
| align="center" rowspan=2 style="background:#f0f0f0;"|'''lane'''
** 3: 5 {{ul}} 2 {{um}} streptavidin (10 pmol), 2 {{ul}} loading dye (+ 5 {{ul}} water)
| align="center" colspan=2 style="background:#f0f0f0;"|'''streptavidin'''
** 4: 5 {{ul}} 1 {{um}} streptavidin (5 pmol), 2 {{ul}} loading dye (+ 5 {{ul}} water)
| align="center" rowspan=2 style="background:#f0f0f0;"|'''water''' ({{ul}})
** 5: 5 {{ul}} 0.5 {{um}} streptavidin (2.5 pmol), 2 {{ul}} loading dye (+ 5 {{ul}} water)
| align="center" rowspan=2 style="background:#f0f0f0;"|'''10x Tris-gly loading dye''' ({{ul}})
** 6: 2 {{ul}} 0.5 {{um}} streptavidin (1 pmol), 2 {{ul}} loading dye (+ 8 {{ul}} water)
|-
** 7: 1 {{ul}} 0.5 {{um}} streptavidin (500 fmol), 2 {{ul}} loading dye (+ 9 {{ul}} water)
| align="center"|({{ul}})|| align="center"|(fmols)
** 8: 1 {{ul}} 0.25 {{um}} streptavidin (250 fmol), 2 {{ul}} loading dye (+ 9 {{ul}} water)
|-
** 9: 1 {{ul}} 0.10 {{um}} streptavidin (100 fmol), 2 {{ul}} loading dye (+ 9 {{ul}} water)
| 1||10 {{ul}} 4 {{um}}||40,000||0||2
** 10: 1 {{ul}} 0.05 {{um}} streptavidin (50 fmol), 2 {{ul}} loading dye (+ 9 {{ul}} water)
|-
| 2||10 {{ul}} 2 {{um}}||20,000||0||2
|-
| 3||5 {{ul}} 2 {{um}}||10,000||5||2
|-
| 4||5 {{ul}} 1 {{um}}||5,000||5||2
|-
| 5||5 {{ul}} 0.5 {{um}}||2,500||5||2
|-
| 6||2 {{ul}} 0.5 {{um}}||1,000||8||2
|-
| 7||1 {{ul}} 0.5 {{um}}||500||9||2
|-
| 8||2.5 {{ul}} 0.1 {{um}}||250||7.5||2
|-
| 9||1 {{ul}} 0.1 {{um}}||100||9||2
|-
| 10||1 {{ul}} 0.05 {{um}}||50||9||2
|}

Revision as of 12:57, 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.
  • 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
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