Team:KIT-Kyoto/Notebook-week5

From 2012.igem.org

(Difference between revisions)
m
m
Line 99: Line 99:
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>
-
,img src="http://2012.igem.org/wiki/images/b/b0/0904kit.png" width="500" height="300">
+
<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="http://2012.igem.org/wiki/images/b/b0/0904kit.png" width="500" height="300">
<br><br>
<br><br>
<h2>September 5th</h2>
<h2>September 5th</h2>
Line 105: Line 132:
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="http://2012.igem.org/wiki/images/3/3a/0905akit.png" width="500" height="300">
<img src="http://2012.igem.org/wiki/images/3/3a/0905akit.png" width="500" height="300">
<br><br>
<br><br>

Revision as of 01:50, 23 September 2012






September 4th


The female-virgin flies (yw) and male flies (yw) were collected for mating with the microinjected males and females.

9/3 GAL440uL
M buffer(TOYOBO)5uL
XbaⅠ(TOYOBO)0.5uL
SpeⅠ(TOYOBO)0.5uL
dH2O4uL
Total50uL


each DNA40uL
3 buffer(NEB)5uL
BglⅡ(NEB)0.5uL
dH2O4.5uL
Total50uL


9/3 GAL423uL
M buffer(TOYOBO)10uL
XbaⅠ(TOYOBO)1uL
SpeⅠ(TOYOBO)1uL
CIP0.5uL
dH2O64.5uL
Total100uL




September 5th


The female-virgin flies (yw) and male flies (yw) were collected and separately kept at 25℃ for mating with the microinjected males and females.

EGFPpSB1C3
DNA template40uL23uL
3 buffer5uL5uL
BglⅡ0.5uL0.5uL
dH2O4.5uL21.5uL
Total50uL50uL






September 6th


The female-virgin flies (yw) and male flies (yw) were collected and separately kept at 25℃ for mating with the microinjected males and females.



September 7th


The eclosed male and virgin female flies from microinjected embryos were collected and kept at 25℃ for maturation.







September 8th


The 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 9th


The 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 10th


The 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.