Team:HokkaidoU Japan/Notebook/August17

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|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/''Hin''d III]  
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===→[[Team:HokkaidoU_Japan/Protocols|Gel Extraction]]===
===→[[Team:HokkaidoU_Japan/Protocols|Gel Extraction]]===
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==Gel Extraction==
 
[[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  Lambda/''Hin''d III]
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|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png  λ/''Hin''d III]
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* Band of RBS isn`t visible
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* 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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Because we were planing to use pSB1C3 we made LB Medium with chloramphenicol
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 12.5 g LB-broth and 7.5 g of Agar 7.5 g to 500 mL to autoclaved distiled water, autoclave
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* Added 500 uL of Chloramphenicol (34mg/mL)
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* Added 500 uL of Chloramphenicol (34 mg/mL)
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* Poured it to 25 plates, 20mL per plate
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* Poured it to 25 plates, 20　mL per plate

Latest revision as of 07:36, 27 October 2010

Notebook

Gel Extraction

Electrophoresis

Electrophoresis After Restriction Enzyme Digestion

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

Electrophoresis to measure consentration after 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
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