IGEM:Harvard/2006/DNA nanostructures/Notebook/2006-7-12: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
(Protein-staining control experiment) |
No edit summary |
||
| Line 14: | Line 14: | ||
Protein-staining control experiment | Protein-staining control experiment | ||
* goal: to load varying concentrations of thrombin onto a gel and stain with Coomassie blue in an attempt to see protein | * goal: to load varying concentrations of thrombin onto a gel and stain with Coomassie blue in an attempt to see protein | ||
* total "reaction" volume: 11 {{ul}} | |||
{| {{table}} | {| {{table}} | ||
| align="center" style="background:#f0f0f0;"|'''Tube''' | | align="center" style="background:#f0f0f0;"|'''Tube''' | ||
| Line 22: | Line 23: | ||
| align="center" style="background:#f0f0f0;"|'''10x loading dye''' ({{ul}}) | | align="center" style="background:#f0f0f0;"|'''10x loading dye''' ({{ul}}) | ||
|- | |- | ||
| 1||||0.65||0.5||8.5|| | | 1||||0.65||0.5||8.5||2.0 | ||
|- | |- | ||
| 2||||1.3||1.0||8.0|| | | 2||||1.3||1.0||8.0||2.0 | ||
|- | |- | ||
| 3||||2.6||2.0||7.0|| | | 3||||2.6||2.0||7.0||2.0 | ||
|- | |- | ||
| 4||||3.9||3.0||6.0|| | | 4||||3.9||3.0||6.0||2.0 | ||
|- | |- | ||
| 5||||5.2||4.0||5.0|| | | 5||||5.2||4.0||5.0||2.0 | ||
|} | |} | ||
* all reagents were mixed in individual 0.2 mL PCR tubes | * all reagents were mixed in individual 0.2 mL PCR tubes | ||
Revision as of 08:34, 12 July 2006
Results from 7/11 Thrombin-Binding Experiment
- gels removed at 10:00 am: appear to have run off the gel (very hard to tell since they were so faint to begin with)
- removed gels from plastic
- cut bottom of gel where it sticks out of narrow hole at the bottom
- pried two plastic sides apart (gel now stuck to one side)
- loosen top edge by sliding flat tool under it a bit.
- make sure the very bottom is completely cut
- run it under water over the a plastic tupperware container - move it back and forth left to right - eventually the gel should come free into the container.
- hold the gel in the container while you pour the water out.
- stained with Coomassie blue (in brown bottle in first 4degree fridge)
- pour enough coomassie blue over it to cover the gel.
- put the lid on the tupperware and put it on the rocker for about 20 minutes.
Protein-staining control experiment
- goal: to load varying concentrations of thrombin onto a gel and stain with Coomassie blue in an attempt to see protein
- total "reaction" volume: 11 μL
| Tube | Lane | Thrombin mass (μg) | 2 μM thrombin (μL) (2 μM = 1.299 μg/μL) |
water (μL) | 10x loading dye (μL) |
| 1 | 0.65 | 0.5 | 8.5 | 2.0 | |
| 2 | 1.3 | 1.0 | 8.0 | 2.0 | |
| 3 | 2.6 | 2.0 | 7.0 | 2.0 | |
| 4 | 3.9 | 3.0 | 6.0 | 2.0 | |
| 5 | 5.2 | 4.0 | 5.0 | 2.0 |
- all reagents were mixed in individual 0.2 mL PCR tubes
