Team:HokkaidoU Japan/Notebook/August17

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(電気泳動)
 
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{{Template:HokkaidoU_Japan}}<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/August16|August 16]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/August18|August 18]]</div></div>
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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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!Lane
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|'''Lane'''
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!DNA
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|'''DNA'''
|-
|-
|1
|1
-
|空き
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|Empty
|-
|-
|2
|2
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|λ/''Hin''d III
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|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''d III]
|-
|-
|3
|3
-
|空き
+
|Empty
|-
|-
|4
|4
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|-
|-
|14
|14
-
|空き
+
|Empty
|-
|-
|15
|15
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|-
|-
|16
|16
-
|空き
+
|Empty
|-
|-
|17
|17
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|空き
+
|Empty
|}
|}
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===→ゲル抽出===
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===→[[Team:HokkaidoU_Japan/Protocols|Gel Extraction]]===
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==電気泳動==
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[[Image:HokkaidoU Japan 20100817b.jpg‎|200px|right|thumb|Electrophoresis to measure consentration after gel extraction]]
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[[Image:HokkaidoU Japan 20100817b.jpg‎|200px|right|thumb|]]
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* After ectraction DNA was electrophoresed to check consentration
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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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* 2 uL of 6x SB was added to DNA solution of 10 uL
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* DNA solution 10 uL に6x SB 2 uL
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{| class="protocol"
{| class="protocol"
|-
|-
-
|レーン
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|'''Lane'''
-
|
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|'''DNA'''
|-
|-
|1
|1
-
|空き
+
|Empty
|-
|-
|2
|2
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|λ/''Hin''d III
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|[https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png  λ/''Hin''d III]
|-
|-
|3
|3
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|-
|-
|8
|8
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|空き
+
|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
 +
→will try again tomorrow
<div style="clear:both"></div>
<div style="clear:both"></div>
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=Chloramphenicol LB培地の作製=
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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
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=Making of Chloramphenicol LB Medium=
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* 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
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Because we were planing to use pSB1C3 we made LB Medium with chloramphenicol
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* 500 uLのChloramphenicol (34mg/mL) を加えた
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* 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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* 25枚のプレートに20 mLずつ分注した
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* 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

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