Difference between revisions of "20.109(S18): Prep notes for M1"
From Course Wiki
Noreen Lyell (Talk | contribs) |
Noreen Lyell (Talk | contribs) (→M2D2) |
||
| Line 22: | Line 22: | ||
** | ** | ||
*250 μL slurry | *250 μL slurry | ||
| + | *7 mL 1X PBS | ||
| + | *Y μL 1 M MgCl<sub>2</sub> | ||
| + | *12 μL DNase | ||
| + | *1.7 mL 1X PBS containing 10 mM imidazole | ||
| + | *3.5 mL 1X PBS containing 250 mM imidazole | ||
| + | |||
| + | ==M2D3== | ||
| + | '''on front bench (per team):''' | ||
| + | |||
| + | *20 μL Laemmli buffer | ||
| + | * | ||
Revision as of 21:26, 31 January 2018
M1D1
- restriction enzymes and buffers
M1D2
prior to laboratory:
- prepare 1% agarose gels, 1 per 2 teams
- subculture BL21(DE3) pRSETb_FKBP12 1:10 into 50 mL LB containing 100μg / mL amp and 34 μg / mL cam, 1 per team
on front bench (per team):
- X μL 6x loading dye
- Y μL 1 M IPTG
M2D2
on front bench (per team):
- lysis buffer reagents
- 250 μL slurry
- 7 mL 1X PBS
- Y μL 1 M MgCl2
- 12 μL DNase
- 1.7 mL 1X PBS containing 10 mM imidazole
- 3.5 mL 1X PBS containing 250 mM imidazole
M2D3
on front bench (per team):
- 20 μL Laemmli buffer
