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
+
* Gel extraction (gel weight: 220 mg)
 +
* Final volume after extraction was50 uL(500 ng/uL)
===Ligation===
===Ligation===
 +
{|style="text-align:center;" class="protocol"
{|style="text-align:center;" class="protocol"
|-
|-
-
|style="border-right:1px solid #000; border-bottom:1px solid #000; width:100px"|
+
|style="border-right:1px solid #996; border-bottom:1px solid #996; width:100px"|
-
|style="border-bottom:1px solid #000;"|なし
+
|style="border-bottom:1px solid #996;"|Empty
-
|style="border-bottom:1px solid #000;"|I
+
|style="border-bottom:1px solid #996;"|I
-
|style="border-bottom:1px solid #000;"|M
+
|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
|-
|-
-
|style="border-right:1px solid #000;"|ligation solution I
+
|style="border-right:1px solid #996;"|ligation solution I
|-
|-
|2 uL
|2 uL
|-
|-
|-
|-
-
|style="border-right:1px solid #000;"|Mighty Mix
+
|style="border-right:1px solid #996;"|Mighty Mix
|-
|-
|-
|-
|2 uL
|2 uL
|-
|-
-
|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
|-
|-
-
|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
-
|style="border-top:1px solid #000;"|4.4 uL
+
|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

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