Team:KIT-Kyoto/Notebook-week6

From 2012.igem.org

(Difference between revisions)
Line 341: Line 341:
<img src="https://static.igem.org/mediawiki/2012/2/26/0917kit.png" width="500" height="300">
<img src="https://static.igem.org/mediawiki/2012/2/26/0917kit.png" width="500" height="300">
 +
<div align="right"><a href="https://2012.igem.org/Team:KIT-Kyoto/Notebook-week7">>>>>>>>>>WEEK7</a></div>
</div>
</div>
</div>
</div>

Revision as of 09:28, 24 September 2012

September 11th


*TNFA and API2

The female-virgin flies (yw) and male flies (yw) were again collected and separately kept at 25℃.

*Parts

　pSB1C3-UAS-1, -2, -3, -4　　5uL　
　3buffer(NEB)　　2uL　
　BglⅡ　　0.5uL　
　dH2O　　12.5uL　
　Total　　20uL　




　GAL4　　40uL　
　4 buffer(NEB)　　5uL　
　XbaⅠ(NEB)　　0.5uL　
　SpeⅠ(NEB)　　0.5uL　
　CIP　　0.5uL　
　100×BSA　　0.5uL　
　dH2O　　3uL　
　Total　　40uL　




　1uL　
　2.5uL　
　2.5uL　
　Ligation high　　6uL　
　Total　　12uL　


　2uL　
　2.5uL　
　1.5uL　
　Ligation high　　6uL　
　Total　　12uL　


September 12th


*TNFA and API2

The female-virgin flies (yw) and male flies (yw) were again collected and separately kept at 25℃.

*Parts

　pSB1C3-UAS-1, -2　　40uL　
　3buffer(NEB)　　5uL　
　SpeⅠ　　1uL　
　100×BSA　　0.5uL　
　dH2O　　3.5uL　
　Total　　50uL　




　1uL　
　2uL　
　EGFP or LacZ　　2uL　
　Ligation high　　2.5uL　
　Total　　7.5uL　


　2uL　
　2uL　
　2uL　
　Ligation high　　6uL　
　Total　　12uL　


September 13th


*TNFA and API2

The female-virgin flies (yw) and male flies (yw) were again collected and separately kept at 25℃.

*Parts

　DNA sample　　1uL　
　4 buffer(NEB)　　0.5uL　
　EcoRⅠ-HF(NEB)　　0.2uL　
　SpeⅠ(NEB)　　0.2uL　
　100×BSA　　0.05uL　
　dH2O　　3.25uL　
　Total　　5uL　


　DNA sample　　1uL　
　3 buffer(NEB)　　0.5uL　
　BglⅡ　　0.2uL　
　dH2O　　3.3uL　
　Total　　5uL　


　1uL　
　1.5uL　
　dH2O　　2.5uL　
　Ligation high　　2.5uL　
　Total　　7.5uL　


　2.5uL　
　2.5uL　
　2uL　
　Ligation high　　7uL　
　Total　　14uL　


　1uL　
　2uL　
　dH2O　　2uL　
　Ligation high　　5uL　
　Total　　10uL　


September 14th


*TNFA and API2

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 female-virgin flies (yw) and male flies (yw) were again collected and separately kept at 25℃.

*Parts

　DNA sample　　1uL　
　4 buffer(NEB)　　1uL　
　EcoRⅠ-HF(NEB)　　0.2uL　
　SpeⅠ(NEB)　　0.2uL　
　100×BSA　　0.1uL　
　dH2O　　7.5uL　
　Total　　10uL　


　2uL　
　3uL　
　1uL(HS) or 2.5uL(Act5c)　
　Ligation high　　6uL(HS) or 7.5uL(Act5c)　
　Total　　12uL(HS) or 15uL(Act5c)　


September 15th


*TNFA and API2

The female-virgin flies (yw) and male flies (yw) were again collected and separately kept at 25℃.

*Parts

　DNA sample　　1uL　
　4 buffer(NEB)　　1uL　
　EcoRⅠ-HF(NEB)　　0.2uL　
　SpeⅠ(NEB)　　0.2uL　
　100×BSA　　0.1uL　
　dH2O　　7.5uL　
　Total　　10uL　




September 16th


*TNFA and API2

The adult male and virgin female flies from microinjected embryos were mated with yw virgin female flies and yw male flies, respectively. In total 83 flies from microinjected embryos were mated with yw flies. Mated flies were transferred to the new food vials in every three days to lay eggs as much as possible.

September 17th


*TNFA and API2

The progeny flies were inspected by dissecting microscope to look for the successfully transformed w+ red eye flies (red eye screening). However, no red eye fly was found.

*Parts

　DNA sample　　1uL　
　4 buffer(NEB)　　1uL　
　EcoRⅠ-HF(NEB)　　0.2uL　
　SpeⅠ(NEB)　　0.2uL　
　100×BSA　　0.1uL　
　dH2O　　7.5uL　
　Total　　10uL　


Retrieved from "http://2012.igem.org/Team:KIT-Kyoto/Notebook-week6"