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Team:TMU-Tokyo/Notebook/Assay 3 Protocol and Result
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<b>■Assay3 Protocol</b><Br> | <b>■Assay3 Protocol</b><Br> | ||
| - | <Br>Extraction of the crude enzyme solution<Br> | + | <Br>Ⅰ.Extraction of the crude enzyme solution<Br> |
| - | <Br> | + | <Br> |
| - | + | <b>1.</b>The cells were cultured at 30 ℃ for 18 hours in medium<Br> | |
| - | + | <b>2.</b>Centrifuged 10000 × g 5min<Br> | |
| - | + | <b>3.</b>Remove the supernatant<Br> | |
| - | E. coli disrupted by sonicator in buffer of 50mM Tris-HCl pH7.2 .<Br> | + | <b>4.</b>Wash the fungus<Br> |
| - | Centrifuged | + | Add dw and Centrifuged 10000 × g 5min<Br> |
| + | <b>5.</b>Remove the supernatant<Br> | ||
| + | <b>6.</b>E. coli disrupted by sonicator in buffer of 50mM Tris-HCl pH7.2 .<Br> | ||
| + | <b>7.</b>Centrifuged 11000 × g at 4 ℃ so that the enzyme is not deactivated, the supernatant was harvested as crude enzyme solution (If you want to save is stored at 4 ℃ in a cold room)<Br> | ||
Revision as of 00:04, 27 September 2012
■Experiment
1st week (8.13 - 8.19)
2nd week (8.20 - 8.26)
3rd week (8.27 - 9. 2)
4th week (9. 3 - 9. 9)
5th week (9.10 - 9.16)
6th week (9.17 - 9.23)
7th week (9.24 - 9.30)
■Protocols
■Assay
Device1 Assay
Device2 Assay
Device3 Assay
Assay 3
■Assay3 Protocol
Ⅰ.Extraction of the crude enzyme solution
1.The cells were cultured at 30 ℃ for 18 hours in medium
2.Centrifuged 10000 × g 5min
3.Remove the supernatant
4.Wash the fungus
Add dw and Centrifuged 10000 × g 5min
5.Remove the supernatant
6.E. coli disrupted by sonicator in buffer of 50mM Tris-HCl pH7.2 .
7.Centrifuged 11000 × g at 4 ℃ so that the enzyme is not deactivated, the supernatant was harvested as crude enzyme solution (If you want to save is stored at 4 ℃ in a cold room)
■Assay3 Result
BBa_K749015
30min 50μl 229371.2 80.88
10min 20μl 248294.2 83.9823
10min No enzym 243418.6 84.37
WT
10min 20μl 232791.6 83.691
