Team:KIT-Kyoto/Notebook-week11

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[[Team:KIT-Kyoto|Home]] > [[Team:KIT-Kyoto/Note|Notebook]] > [[Team:KIT-Kyoto/Notebook|Lab Note]] > [[Team:KIT-Kyoto/Notebook-week8|Week8]]</td></tr></table>
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【時間】 9:00~18:00<BR>
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<BR>
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【実験担当】岩城,福山,竹内,臼井,足立<BR>
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<BR>
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【実験名】<BR>
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(1)pSB6及びpsB1(非パーツ)のプレカルチャー<BR>
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(2)pGVBpttの電気泳動<BR>
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(3)psB3K3(J04450)のトランスフォーメーション<BR>
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<BR>
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【実験目的】<BR>
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(1)pSB6A1(K121013)及びpsB1(非パーツ)のプレカルチャー<BR>
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(2)pGVBpttが正しく制限酵素処理されているかどうかを確認するため<BR>
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(3)psB3K3(J04450)のトランスフォーメーション<BR>
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<BR>
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【実験内容】<BR>
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<BR>
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(1)プレカルチャー<BR>
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:DH5α-pSB6を液体LB培地(+amp)2mLで培養<BR>
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:DH5α-pSB1 を液体LB培地(+amp)2mLで培養<BR>
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(2)pGVBpttの電気泳動<BR>
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:〈組成〉<BR>
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::pGVBppt 5μL<BR>
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::KpmI 1μL <BR>
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::HindⅢ 1μL <BR>
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::Buffer(M) 1μL <BR>
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::H2O 11μL <BR>
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::計  20μL  <BR>
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<BR>
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:これを1h,37℃でインキュベート<BR>
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:        ↓<BR>
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:Loading Buffer 1μLを加えたものと、マーカー(1kbp radder)を<BR>
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:1% Agarose Gel のコームに流し込んだ<BR>
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:        ↓100V,30min泳動<BR>
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:        ↓EtBr(10mg/mL)で20min染色<BR>
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:泳動結果を写真で撮影した<BR>
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(3)psB3K3にSOC 0.9mLを加えDH5αにトランスフォーメーションした<BR>
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:             ↓<BR>
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:  37℃でインキュベート(Overnight)<BR>
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<BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR><BR>
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Latest revision as of 01:20, 21 October 2010



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