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Team:Tsinghua/Notebook/14 July 2010
From 2010.igem.org
(Difference between revisions)
(→Module I, DT and Fan's part:) |
(→Module I, DT and Fan's part:) |
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eGFP: | eGFP: | ||
| - | H2O 23.5μl | + | H2O 23.5μl |
10×buffer 5μl | 10×buffer 5μl | ||
| - | eGFP 20μl | + | eGFP 20μl |
HindIII 1.5μl | HindIII 1.5μl | ||
| - | Total 50μl | + | Total 50μl |
mCherry: | mCherry: | ||
| - | H2O 11.5μl | + | H2O 11.5μl |
10×buffer 5μl | 10×buffer 5μl | ||
mCherry 32μl | mCherry 32μl | ||
HindIII 1.5μl | HindIII 1.5μl | ||
| - | Total 50μl | + | Total 50μl |
Kan: | Kan: | ||
| - | H2O 31μl | + | H2O 31μl |
10×buffer 5μl | 10×buffer 5μl | ||
| - | Kan 12.5μl | + | Kan 12.5μl |
| - | KpnI 1.5μl | + | KpnI 1.5μl |
| - | Total 50μl | + | Total 50μl |
Chlr: | Chlr: | ||
| - | H2O 31.5μl | + | H2O 31.5μl |
10×buffer 5μl | 10×buffer 5μl | ||
| - | Chlr 12μl | + | Chlr 12μl |
| - | KpnI 1.5μl | + | KpnI 1.5μl |
| - | Total 50μl | + | Total 50μl |
Revision as of 09:11, 3 September 2010
Module I, DT and Fan's part:
Measure the concentration of the PCR products.
Gene Concentration(μg/ml) OD 260/280 eGFP 95.9 1.73 mCherry 61.2 1.74 Kan 160.5 1.67 Chlr 167.5 1.74
Digest the four fragments with follow system at 37℃ for 10 hours.
Digestion system:
eGFP:
H2O 23.5μl 10×buffer 5μl eGFP 20μl HindIII 1.5μl Total 50μl
mCherry:
H2O 11.5μl 10×buffer 5μl mCherry 32μl HindIII 1.5μl Total 50μl
Kan:
H2O 31μl 10×buffer 5μl Kan 12.5μl KpnI 1.5μl Total 50μl
Chlr:
H2O 31.5μl 10×buffer 5μl Chlr 12μl KpnI 1.5μl Total 50μl
Productions purify and measure the concentration.
Gene Concentration(μg/ml) OD 260/280 eGFP 38.4 1.66 mCherry 28.8 1.86 Kan 43.7 2.11 Chlr 31.9 1.77
