Biomolecular Breadboards:Protocols:Crude Extract Prep: Difference between revisions
From OpenWetWare
Jump to navigationJump to search
Clare Hayes (talk | contribs) m (New page: ==Protocol for ''E. Coli'' 3x Crude Extract Preparation== Time needed: 3 days ===Materials=== 2xYT: *2xYT-broth powder (MP biomedicals) *Cat #: 3012-032 Bacto-Agar: *Bacto-agar (BD) *Prod...) |
Clare Hayes (talk | contribs) mNo edit summary |
||
Line 1: | Line 1: | ||
==Protocol for ''E. Coli'' | ==Protocol for 3x Crude Extract Preparation from ''E. Coli''== | ||
Time needed: 3 days | Time needed: 3 days | ||
===Materials=== | ===Materials=== |
Revision as of 18:46, 4 June 2012
Protocol for 3x Crude Extract Preparation from E. Coli
Time needed: 3 days
Materials
2xYT:
- 2xYT-broth powder (MP biomedicals)
- Cat #: 3012-032
Bacto-Agar:
- Bacto-agar (BD)
- Product number: 214010
Bead:
- 0.1mm diameter glass beads (Biospec products)
- Catalog #: 11079101
Bead beater:
- Mini-Beadbeater-1 (Biospec products)
- Catalog #: 3110BX
Bead beating tube:
- Polypropylene microvials (Biospec products)
- Catalog #: 522S
Bead filter:
- Micro bio-spin chromatography columns (Bio-Rad)
- Product number: 732-6204
Centrifuge (big size):
- Allegra 64R high-speed refrigerated benchtop centrifuge (Beckman Coulter)
Centrifuge (ultra size):
- Beckman J2-M1 (Beckman Coulter)
Centrifuge bottle (50ml):
- Polyallomer bottle with screw-on cap (Beckman Coulter)
- Part number: 357003
Centrifuge bottle (500ml):
- Centrifuge bottle assembly polycarbonate (Beckman Coulter)
- Part number: 355607
Chloramphenicol:
- Chloramphenicol powder (Sigma-Aldrich)
- Product number: C1919
Cuvette:
- Fisherbrand Disposable Cuvettes
- Cat#: 14-955-127
Dialysis cassette:
- Slide-A-Lyzer Dialysis Cassette, 10MWCO (Thermo Scientific), maximum capacity: 3mL
- Product number: 66380
E. coli strain:
- BL21 Rosetta 2 competent cell (Novagen)
- Product number: 71402
Electrode:
- Electrode, 3-in-1 for Φ 400, 500 series pH meter, 12x 150 mm, Epoxy
- Product number: A57198
Erlenmeyer glass flask:
- KIMAX erlenmeyer glass flask
- Product number: 26500 (4000ml)
Floater:
- Float Buoys: comes with Slide-A-Lyzer Dialysis cassette
Incubator shaker:
- Incubator shaker (New Brunswick)
- Innova 4300
K-glutamate:
- L-Glutamic acid potassium salt monohydrate (Sigma Aldrich)
- Product number: G1149
Liquid nitrogen
Magnetic stirrer:
- Thermolyne Nuova II
- Fisher Scienetific Isotemp Digital Magnetic Stirrer
Mg-glutamate:
- L-Glutamic acid Hemimagnesium salt tetrahydrate (Sigma Aldrich)
- Product number: 49605
PH meter:
- pH meter, Φ 410 pH/mV meter, waterproof, handheld
- Product number: A58734
Potassium phosphate monobasic solution:
- Potassium phosphate monobasic solution (Sigma Aldrich)
- Product number: P8709
Potassium phosphate dibasic solution:
- Potassium phosphate dibasic solution (Sigma Aldrich)
- Product number: P8584
Protein assay:
- Bio-rad Protein Assay Dye Reagent concentrate (Bio-Rad)
- Product number: 500-0006
Spectrometer:
- Genesys 10 uv (Thermo Electron corporation)
Table top centrifuge:
- Microcentrifuge model 5415D (Eppendorf)
Table top vortex:
- Analog vortex mixer (VWR)
- Catalog number: 58816-121
Tris:
- Tris base, white crystals or crystalline powder, molecular biology (Fisher Scientific)
- Catalog number: BP 1 52 1