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WT Hemoglobin Expression
- Expression continued from yesterday.
- In morning, spin down starter cultures (4500rpm for 20min at 4C)
- Resuspend cell pellets in fresh broth and divide evenly among 1L expression flasks
- Incubate at 37C and shake at 165rpm
- When OD(600) reaches 0.6 induce expression by adding 1mL of 0.4M IPTG and 1mL of 40mg/mL hemin in DMF to cultures
- Change shaker temperature to 30C
- 4 hours after inducing, harvest cells in centrifuge (4500rpm, 30min, 4C)
- Resuspend cells in 50mL of 25mM Tris, 50mM NaCl, 1mM EDTA, 0.5mM DTT
- Store in -80C Freezer
- Add data and results here...
This area is for any observations or conclusions that you would like to note.
Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.