Team:HokkaidoU Japan/Notebook/August12

From 2010.igem.org

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** Melted in 30 uL of TE and did electrophoresis
** Melted in 30 uL of TE and did electrophoresis
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=電気泳動２=
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=Electrophoresis ２=
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[[Image:HokkaidoU Japan 20100812b.jpg‎|200px|right|thumb|撮影1回目]]
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[[Image:HokkaidoU Japan 20100812c.jpg‎|200px|right|thumb|撮影2回目]]
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[[Image:HokkaidoU Japan 20100812b.jpg‎|200px|right|thumb|Take 1]]
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mini prepで得たプラスミドとPCRで得たパーツのDNAを電気泳動で確認した<br>
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[[Image:HokkaidoU Japan 20100812c.jpg‎|200px|right|thumb|Take 2 after few minutes]]
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制限酵素処理はしないので、反応系は以下の通り
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Checked plasmids gathered via mini prep and PCR products, by electrophoresis. <br>
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Because we wont do restriction enzyme digestion reaction system is ass follows.
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{| style="text-align:center" class="protocol"
{| style="text-align:center" class="protocol"
|-
|-
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|style="border-bottom:1px solid #000; border-right:1px solid #000;"|
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|style="border-bottom:1px solid #000; border-right:1px solid #000;"|'''Lane'''
|style="border-bottom:1px solid #000;"| １
|style="border-bottom:1px solid #000;"| １
|style="border-bottom:1px solid #000;"| ２
|style="border-bottom:1px solid #000;"| ２
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| 0.4  
| 0.4  
|}
|}
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* マーカーはλ/''Hin''dIIIとpUC119/''Hin''fIを使用
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* Used markers are [https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/''Hin''dIII] and [https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png pUC119/''Hin''fI]
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* レーン１のマーカー（λ/''Hin''dIII）はウェルから流れ出てしまった
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* Marker in lane 1 ([https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png Lambda/''Hin''dIII]) leaked out
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* レーン３（mini prepのサンプル，1-18F）はバンドが確認できなかった（後日PCR）
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* Mini prep sample，1-18F band in lane 3 wasn't visible so we decided PCR it tomorrow
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* レーン４ではplasmidのダイマー，トライマーがよく確認できる（plasmidの電気泳動ではよくあるらしい）
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* In lane 4 there were some visible plasmid dimers and trimer s

Revision as of 06:51, 20 September 2010

Notebook

Electrophoresis 1

Electrophoresis of yesterdays ligation of PCR products
  • Checked the result of yesterdays ligation of PCR products by electrophoresis
  • Added 4 uL of 6x Sample Buffer to make 12 uL in total
  • Used 20uL of EtOh

Mini prep

  • Used cells incubated for 18h in LB broth
    • 1-18F didn't grow well
  • Centrifuge after adding 430 uL of chloroform、collected supernatant
  • Added 2-propanol to 1-18F, but after centrifugation there were no precipitation
    • Melted in 30 uL of TE and did electrophoresis

Electrophoresis ２

Take 1
Take 2 after few minutes

Checked plasmids gathered via mini prep and PCR products, by electrophoresis.


Because we wont do restriction enzyme digestion reaction system is ass follows.

Lane １ ２ ３ ４ ５ ６
Parts 1-18F 2-21H 2-11P 2-24G 1-2M 3-1E
DNA solution 2 uL 2 2 1 1 1
6x Sample Buffer 0.4 uL 0.4 0.4 0.4 0.4 0.4
  • Used markers are Lambda/HindIII and pUC119/HinfI
  • Marker in lane 1 (Lambda/HindIII) leaked out
  • Mini prep sample，1-18F band in lane 3 wasn't visible so we decided PCR it tomorrow
  • In lane 4 there were some visible plasmid dimers and trimer s
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