Team:HokkaidoU Japan/Notebook/August31

From 2010.igem.org

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=pUC119のゲル抽＆Ligation&Transformation=
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=[[Team:HokkaidoU_Japan/Protocols|Gel extraction]] of pUC119, Ligation and [[Team:HokkaidoU_Japan/Protocols|Transformation]]=
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* pUC119 solution 50 uLに 10 uL 6x Sample Bufferを加え，３レーンに流した
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* Added 10 uL 6x Sample Buffer to pUC119 solution 50 uL
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* ゲル抽出(220 mg)
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* Divided among 3 lanes
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* 50 uL（500 ng/uL）になったとする
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* Gel extraction (gel weight: 220 mg)
 +
* Final volume after extraction was50 uL（500 ng/uL）
===Ligation===
===Ligation===
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{|style="text-align:center;" class="protocol"
{|style="text-align:center;" class="protocol"
|-
|-
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|style="border-right:1px solid #000; border-bottom:1px solid #000; width:100px"|
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|style="border-right:1px solid #996; border-bottom:1px solid #996; width:100px"|
-
|style="border-bottom:1px solid #000;"|なし
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|style="border-bottom:1px solid #996;"|Empty
-
|style="border-bottom:1px solid #000;"|I
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|style="border-bottom:1px solid #996;"|I
-
|style="border-bottom:1px solid #000;"|M
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|style="border-bottom:1px solid #996;"|M
|-
|-
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|style="border-right:1px solid #000;"|DNA
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|style="border-right:1px solid #996;"|DNA
|2 uL
|2 uL
|2 uL
|2 uL
|2 uL
|2 uL
|-
|-
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|style="border-right:1px solid #000;"|ligation solution I
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|style="border-right:1px solid #996;"|ligation solution I
|－
|－
|2 uL
|2 uL
|－
|－
|-
|-
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|style="border-right:1px solid #000;"|Mighty Mix
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|style="border-right:1px solid #996;"|Mighty Mix
|－
|－
|－
|－
|2 uL
|2 uL
|-
|-
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|style="border-right:1px solid #000;"|T4 ligase
+
|style="border-right:1px solid #996;"|T4 ligase
|－
|－
|0.4 uL
|0.4 uL
|0.4 uL
|0.4 uL
|-
|-
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|style="border-right:1px solid #000; border-top:1px solid #000;"|Total
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|style="border-right:1px solid #996; border-top:1px solid #996;"|Total
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|style="border-top:1px solid #000;"|2 uL
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|style="border-top:1px solid #996;"|2 uL
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|style="border-top:1px solid #000;"|4.4 uL
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|style="border-top:1px solid #996;"|4.4 uL
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|style="border-top:1px solid #000;"|4.4 uL
+
|style="border-top:1px solid #996;"|4.4 uL
|}
|}
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→Transformation
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→[[Team:HokkaidoU_Japan/Protocols|Transformation]]

Latest revision as of 09:13, 2 October 2010

Notebook

Gel extraction of pUC119, Ligation and Transformation

  • Added 10 uL 6x Sample Buffer to pUC119 solution 50 uL
  • Divided among 3 lanes
  • Gel extraction (gel weight: 220 mg)
  • Final volume after extraction was50 uL（500 ng/uL）

Ligation

Empty I M
DNA 2 uL 2 uL 2 uL
ligation solution I － 2 uL －
Mighty Mix － － 2 uL
T4 ligase － 0.4 uL 0.4 uL
Total 2 uL 4.4 uL 4.4 uL
→Transformation
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