Line 145: | Line 145: | ||
</table> | </table> | ||
+ | <br> | ||
+ | After we finished cultivating ?samples, we took 100μl out of each samples and made the following operation. | ||
+ | 1. Centrifuge with 13000rpm at 24℃ for 2min | ||
+ | 2. Remove the supernatant | ||
+ | 3. Add 50ml of SDS-Buffer | ||
+ | 4. Shake for 1min | ||
+ | 5. Keep at 100℃ for 5min | ||
+ | 6. Put on the ice | ||
+ | 7. Apply 10μl to SDS | ||
+ | 8. Run electrophoresis for 1.5h | ||
+ | 9. Wash out with MiliQ | ||
+ | 10. Shake at 24℃ with 34rpm for 1h | ||
+ | 11. Dye with QuickCBB | ||
+ | The fig. 1 shows the result of SDS-PAGE. | ||
+ | </span> | ||
</html> | </html> |
Revision as of 12:46, 18 October 2016
Team:HokkaidoU Japan
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We conducted SDS-PAGE with (BBa_K2015012+BBa_K2015008/ BBa_K2015012+BBa_K2015009) on (pSB1C3)of E.coli(DH5α). At first, we made the Gel for SDS-PAGE, with the following the Table1. What was the most important was to add TEMED ffinally because
The following Table2 is about preparing the SDS-PAGE.
We conducted SDS-PAGE with (BBa_K2015012+BBa_K2015008/ BBa_K2015012+BBa_K2015009) on (pSB1C3)of E.coli(DH5α). At first, we made the Gel for SDS-PAGE, with the following the Table1. What was the most important was to add TEMED ffinally because
Table. 1. Gel assay |
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The following Table2 is about preparing the SDS-PAGE.
Table. 2. Experimental condition |
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After we finished cultivating ?samples, we took 100μl out of each samples and made the following operation. 1. Centrifuge with 13000rpm at 24℃ for 2min 2. Remove the supernatant 3. Add 50ml of SDS-Buffer 4. Shake for 1min 5. Keep at 100℃ for 5min 6. Put on the ice 7. Apply 10μl to SDS 8. Run electrophoresis for 1.5h 9. Wash out with MiliQ 10. Shake at 24℃ with 34rpm for 1h 11. Dye with QuickCBB The fig. 1 shows the result of SDS-PAGE. |