Difference between revisions of "Part:BBa K797006:Experience"
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This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | This experience page is provided so that any user may enter their experience using this part.<BR>Please enter | ||
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===Applications of BBa_K797006=== | ===Applications of BBa_K797006=== | ||
| + | In order to comfirm the expression of FT protein, we performed western blotting. | ||
| + | |||
| + | [[Image:Kyoto2012_Western-FT.png|thumb|600px|left|fig.1-4 Verification of expression of FT protein in ''E.coli''. The result of Western blotting against FT protein. <br> | ||
| + | Each plasmid shown in Fig.1-3 was transformed into BL21(DE3). Cells were precultured overnight and diluted into fresh SOC medium. IPTG was added when OD600 was approx. 0.5, then cells were incubated for 4h or overnight at 20°C. 100µL of culture was used for SDS-PAGE. | ||
| + | Lane1 : Cell lysate diluted into 100µL of sample buffer, not induced<br> | ||
| + | Lane2 : Cell lysate diluted into 100µL of sample buffer, IPTG induced 4h.<br> | ||
| + | Lane3 : Cell lysate diluted into 100µL of sample buffer, IPTG induced overnight<br> | ||
| + | Lane4 : Cell lysate diluted into 50µL of sample buffer, IPTG induced overnight<br> | ||
| + | Lane5 : Cell lysate diluted into 100µL of sample buffer, not induced<br> | ||
| + | Lane6 : Cell lysate diluted into 100µL of sample buffer, IPTG induced 4h.<br> | ||
| + | Lane7 : Cell lysate diluted into 100µL of sample buffer, IPTG induced overnight<br> | ||
| + | Lane8 : Cell lysate diluted into 50µL of sample buffer, IPTG induced overnight | ||
| + | ]] | ||
| + | <br clear="both" /> | ||
===User Reviews=== | ===User Reviews=== | ||
Latest revision as of 23:22, 26 September 2012
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Applications of BBa_K797006
In order to comfirm the expression of FT protein, we performed western blotting.
fig.1-4 Verification of expression of FT protein in E.coli. The result of Western blotting against FT protein.
Each plasmid shown in Fig.1-3 was transformed into BL21(DE3). Cells were precultured overnight and diluted into fresh SOC medium. IPTG was added when OD600 was approx. 0.5, then cells were incubated for 4h or overnight at 20°C. 100µL of culture was used for SDS-PAGE. Lane1 : Cell lysate diluted into 100µL of sample buffer, not induced
Lane2 : Cell lysate diluted into 100µL of sample buffer, IPTG induced 4h.
Lane3 : Cell lysate diluted into 100µL of sample buffer, IPTG induced overnight
Lane4 : Cell lysate diluted into 50µL of sample buffer, IPTG induced overnight
Lane5 : Cell lysate diluted into 100µL of sample buffer, not induced
Lane6 : Cell lysate diluted into 100µL of sample buffer, IPTG induced 4h.
Lane7 : Cell lysate diluted into 100µL of sample buffer, IPTG induced overnight
Lane8 : Cell lysate diluted into 50µL of sample buffer, IPTG induced overnight
Each plasmid shown in Fig.1-3 was transformed into BL21(DE3). Cells were precultured overnight and diluted into fresh SOC medium. IPTG was added when OD600 was approx. 0.5, then cells were incubated for 4h or overnight at 20°C. 100µL of culture was used for SDS-PAGE. Lane1 : Cell lysate diluted into 100µL of sample buffer, not induced
Lane2 : Cell lysate diluted into 100µL of sample buffer, IPTG induced 4h.
Lane3 : Cell lysate diluted into 100µL of sample buffer, IPTG induced overnight
Lane4 : Cell lysate diluted into 50µL of sample buffer, IPTG induced overnight
Lane5 : Cell lysate diluted into 100µL of sample buffer, not induced
Lane6 : Cell lysate diluted into 100µL of sample buffer, IPTG induced 4h.
Lane7 : Cell lysate diluted into 100µL of sample buffer, IPTG induced overnight
Lane8 : Cell lysate diluted into 50µL of sample buffer, IPTG induced overnight
User Reviews
UNIQ8115a9b4fcdcabae-partinfo-00000000-QINU UNIQ8115a9b4fcdcabae-partinfo-00000001-QINU