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Team:KIT-Kyoto/Notebook-week5
From 2012.igem.org
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The female-virgin flies (yw) and male flies (yw) were collected for mating with the microinjected males and females. | The female-virgin flies (yw) and male flies (yw) were collected for mating with the microinjected males and females. | ||
<br><br> | <br><br> | ||
| - | + | <Table Border Cellspacing="0"> | |
| + | <Tr><Td>9/3 GAL4</Td><Td>40uL</Td></Tr> | ||
| + | <tr><td>M buffer(TOYOBO)</td><td>5uL</td></tr> | ||
| + | <Tr><Td>XbaⅠ(TOYOBO)</Td><Td>0.5uL</Td></Tr> | ||
| + | <tr><td>SpeⅠ(TOYOBO)</td><td>0.5uL</td></tr><tr> | ||
| + | <td>dH<sub>2</sub>O</td><td>4uL</td></tr> | ||
| + | <td>Total<td>50uL</td></tr> | ||
| + | </Table> | ||
| + | <br><BR> | ||
| + | <Table Border Cellspacing="0"> | ||
| + | <Tr><Td>each DNA</Td><Td>40uL</Td></Tr> | ||
| + | <tr><td>3 buffer(NEB)</td><td>5uL</td></tr> | ||
| + | <Tr><Td>BglⅡ(NEB)</Td><Td>0.5uL</Td></Tr> | ||
| + | <td>dH<sub>2</sub>O</td><td>4.5uL</td></tr> | ||
| + | <td>Total<td>50uL</td></tr> | ||
| + | </Table> | ||
| + | <br><BR> | ||
| + | <Table Border Cellspacing="0"> | ||
| + | <Tr><Td>9/3 GAL4</Td><Td>23uL</Td></Tr> | ||
| + | <tr><td>M buffer(TOYOBO)</td><td>10uL</td></tr> | ||
| + | <Tr><Td>XbaⅠ(TOYOBO)</Td><Td>1uL</Td></Tr> | ||
| + | <Tr><Td>SpeⅠ(TOYOBO)</Td><Td>1uL</Td></Tr> | ||
| + | <Tr><Td>CIP</Td><Td>0.5uL</Td></Tr> | ||
| + | <td>dH<sub>2</sub>O</td><td>64.5uL</td></tr> | ||
| + | <td>Total<td>100uL</td></tr> | ||
| + | </Table> | ||
| + | <br><BR> | ||
| + | <img src="https://static.igem.org/mediawiki/2012/b/b0/0904kit.png" width="500" height="300"> | ||
<br><br> | <br><br> | ||
<h2>September 5th</h2> | <h2>September 5th</h2> | ||
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The female-virgin flies (yw) and male flies (yw) were collected and separately kept at 25℃ for mating with the microinjected males and females. | The female-virgin flies (yw) and male flies (yw) were collected and separately kept at 25℃ for mating with the microinjected males and females. | ||
<br><br> | <br><br> | ||
| + | <Table Border Cellspacing="0"> | ||
| + | <Tr><Td></Td><Td>EGFP</Td><Td>pSB1C3</Td></Tr> | ||
| + | <tr><td>DNA template</td><td>40uL</td><Td>23uL</Td></tr> | ||
| + | <Tr><Td>3 buffer</Td><Td>5uL</Td><Td>5uL</Td></Tr> | ||
| + | <tr><td>BglⅡ</td><td>0.5uL</td><Td>0.5uL</Td></tr><tr> | ||
| + | <td>dH<sub>2</sub>O</td><td>4.5uL</td><Td>21.5uL</Td></tr> | ||
| + | <td>Total<td>50uL</td><Td>50uL</Td></tr> | ||
| + | </Table> | ||
| + | <br><BR> | ||
<img src="https://static.igem.org/mediawiki/2012/3/3a/0905akit.png" width="500" height="300"> | <img src="https://static.igem.org/mediawiki/2012/3/3a/0905akit.png" width="500" height="300"> | ||
<br><br> | <br><br> | ||
Revision as of 01:50, 23 September 2012
September 4thThe female-virgin flies (yw) and male flies (yw) were collected for mating with the microinjected males and females.
September 5thThe female-virgin flies (yw) and male flies (yw) were collected and separately kept at 25℃ for mating with the microinjected males and females.
September 6thThe female-virgin flies (yw) and male flies (yw) were collected and separately kept at 25℃ for mating with the microinjected males and females. 
September 7thThe eclosed male and virgin female flies from microinjected embryos were collected and kept at 25℃ for maturation. 
September 8thThe adult male and virgin female flies from microinjected embryos were mated with yw virgin female flies and yw male flies, respectively. Mated flies were transferred to the new food vials in every three days to lay eggs as much as possible. The eclosed male and virgin female flies from microinjected embryos were again collected and kept at 25℃ for maturation. The female-virgin flies (yw) and male flies (yw) were collected and separately kept at 25℃. September 9thThe eclosed male and virgin female flies from microinjected embryos were again collected and kept at 25℃ for maturation. In total 83 flies were eclosed from the microinjected embryos. The calculated viability for the microinjected flies was 12.0%. The female-virgin flies (yw) and male flies (yw) were again collected and separately kept at 25℃. 
September 10thThe adult male and virgin female flies from microinjected embryos were mated with yw virgin female flies and yw male flies, respectively. Mated flies were transferred to the new food vials in every three days to lay eggs as much as possible. 
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