Team:HokkaidoU Japan/Notebook/August17
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- | |[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png | + | |[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''d III] |
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[[Image:HokkaidoU Japan 20100817b.jpg|200px|right|thumb|Electrophoresis to measure consentration after gel extraction]] | [[Image:HokkaidoU Japan 20100817b.jpg|200px|right|thumb|Electrophoresis to measure consentration after gel extraction]] | ||
* After ectraction DNA was electrophoresed to check consentration | * After ectraction DNA was electrophoresed to check consentration | ||
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- | |[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png | + | |[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''d III] |
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- | * Band of RBS isn | + | * Band of RBS isn't visible |
** This Might due to loss durring extraction | ** This Might due to loss durring extraction | ||
→will try again tomorrow | →will try again tomorrow | ||
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<div style="clear:both"></div> | <div style="clear:both"></div> | ||
- | =Chloramphenicol | + | =Making of Chloramphenicol LB Medium= |
- | + | Because we were planing to use pSB1C3 we made LB Medium with chloramphenicol | |
- | * LB-broth | + | * Added 12.5 g LB-broth and 7.5 g of Agar 7.5 g to 500 mL to autoclaved distiled water, autoclave |
- | * 500 | + | * Added 500 uL of Chloramphenicol (34 mg/mL) |
- | * | + | * Poured it to 25 plates, 20 mL per plate |
Latest revision as of 07:36, 27 October 2010
Gel Extraction
Electrophoresis
20 uL of DNA digested yesterday was electrophoresed with 4 ul of 6x SB. All acording to the table bellow.
Lane | DNA |
1 | Empty |
2 | λ/Hind III |
3 | Empty |
4 | pSB1C3/E, P |
5 | pSB1C3/E, P |
6 | Heat Shock Promotor/E, S |
7 | Heat Shock Promotor/E, S |
8 | RBS/X, P |
9 | RBS/X, P |
10 | RFP/E, S |
11 | RFP/E, S |
12 | double Terminator/E, S |
13 | double Terminator/E, S |
14 | Empty |
15 | λ/Hind III |
16 | Empty |
17 | Empty |
→Gel Extraction
- After ectraction DNA was electrophoresed to check consentration
- 2 uL of 6x SB was added to DNA solution of 10 uL
Lane | DNA |
1 | Empty |
2 | λ/Hind III |
3 | Vector |
4 | Heat shock promotor |
5 | RBS |
6 | RFP |
7 | double terminator |
8 | Empty |
- Band of RBS isn't visible
- This Might due to loss durring extraction
→will try again tomorrow
Making of Chloramphenicol LB Medium
Because we were planing to use pSB1C3 we made LB Medium with chloramphenicol
- Added 12.5 g LB-broth and 7.5 g of Agar 7.5 g to 500 mL to autoclaved distiled water, autoclave
- Added 500 uL of Chloramphenicol (34 mg/mL)
- Poured it to 25 plates, 20 mL per plate