Team:HokkaidoU Japan/Notebook/August17

From 2010.igem.org

(Difference between revisions)
(→電気泳動)
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=ゲル抽出=
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=Gel Extraction=
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==電気泳動==
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==Electrophoresis==
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[[Image:HokkaidoU Japan 20100817a.JPG‎|200px|right|thumb|]]
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前日に制限酵素処理した20 uLのDNA solutionに4 uLの6x SBを加え，それぞれ2レーンに流した
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[[Image:HokkaidoU Japan 20100817a.JPG‎|200px|right|thumb|Electrophoresis After Restriction Enzyme Digestion]]
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20 uL of DNA digested yesterday was electrophoresed with 4 ul of 6x SB. All acording to the table bellow.
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{|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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|空き
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|Empty
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|4
|4
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|14
|14
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|空き
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|Empty
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|16
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|空き
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|Empty
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|17
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|空き
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|Empty
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===→ゲル抽出===
===→ゲル抽出===
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==電気泳動==
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==Gel Extraction==
[[Image:HokkaidoU Japan 20100817b.jpg‎|200px|right|thumb|]]
[[Image:HokkaidoU Japan 20100817b.jpg‎|200px|right|thumb|]]
* ゲル抽出して得たDNAを電気泳動で確認した．
* ゲル抽出して得たDNAを電気泳動で確認した．

Revision as of 16:05, 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

→ゲル抽出

Gel Extraction

HokkaidoU Japan 20100817b.jpg
  • ゲル抽出して得たDNAを電気泳動で確認した．
  • DNA solution 10 uL に6x SB 2 uL
Lane DNA
1 空き
2 λ/Hind III
3 Vector
4 Heat shock promotor
5 RBS
6 RFP
7 double terminator
8 空き


  • 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

→翌日Digestionからリベンジ

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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