Technique / Cell Biology / In vitro assay and enzyme assay
Temperature control in the T7 RNA polymerase/promoter system. (For protein expre
Temperature control in the T7 RNA polymerase/promoter system. (For protein expre
Why is the temperature control for growing E. coli under the T7 RNA polymerase/promoter system required?
Page 16-6 in "Short Protocols In Molecular Biology 3rd ed. edited by Fred Ausubel et al." explains the procedures for growing E. coli under the T7 RNA polymerase/promoter system :
"...
5) Pick a single E. coli... Inoculate it into 5ml LB/ampcillin/kanamycin medium and grow overnight at *30 degrees C.*.
6) Dilute 1ml 1:40 into LB... and grow several hours at *30 degrees C.* to an OD = 0.4.
7) Induce gene by quikly raising temperature to *42 degrees C.* for *30 min*.
8) Reduce temperature to *37 degrees* C. and grow cells an additional *90 min*... Harvest.
..."
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- Temperature control in the T7 RNA polymerase/promoter system. (For protein expression)?
- Re: Temperature control in the T7 RNA polymerase/promoter system. (For
protein expression)?
- Re: Temperature control in the T7 RNA polymerase/promoter system. (For
protein expression)?
- Re: Temperature control in the T7 RNA polymerase/promoter system. (For
protein expression)?
- Re: Temperature control in the T7 RNA polymerase/promoter system. (For
Last update 15-Nov-2000, Rating of 0 votes.
