Team:HokkaidoU Japan/Notebook/August17

From 2010.igem.org

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==Gel Extraction==
==Gel Extraction==
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[[Image:HokkaidoU Japan 20100817b.jpg‎|200px|right|thumb|]]
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[[Image:HokkaidoU Japan 20100817b.jpg‎|200px|right|thumb|Electrophoresis to measure consentration after gel extraction]]
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* ゲル抽出して得たDNAを電気泳動で確認した．
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* After ectraction DNA was electrophoresed to check consentration
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* DNA solution 10 uL に6x SB 2 uL
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* 2 uL of 6x SB was added to DNA solution of 10 uL
{| class="protocol"
{| class="protocol"
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|1
|1
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|空き
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|Empty
|-
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|2
|2
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|λ/''Hin''d III
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|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png  Lambda/''Hin''d III]
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|3
|3
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|8
|8
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|空き
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|Empty
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* RBSは取れたDNAが少なかったようで，バンドが見られなかった
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* Band of RBS isn`t visible
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→翌日Digestionからリベンジ
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** This Might due to loss durring extraction
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→will try again tomorrow
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Revision as of 16:10, 21 September 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 Lambda/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

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 Lambda/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

Chloramphenicol LB培地の作製

組み上げたパーツはpUC1C3につなぐため，クロラムフェニコール入りのLB寒天培地を作った

  • LB-broth 12.5 g, Agar 7.5 gにDWを加えて500 mLにし，オートクレーブにかけた
  • 500 uLのChloramphenicol (34mg/mL) を加えた
  • 25枚のプレートに20 mLずつ分注した
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