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Team:KIT-Kyoto/Notebook-week11
From 2010.igem.org
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Revision as of 13:59, 6 September 2010
Click me!
Click me!
【時間】 9:00～18:00
【実験担当】岩城,福山,竹内,臼井,足立
【実験名】
(1)pSB6及びpsB1(非パーツ)のプレカルチャー
(2)pGVBpttの電気泳動
(3)psB3K3(J04450)のトランスフォーメーション
【実験目的】
(1)pSB6A1(K121013)及びpsB1(非パーツ)のプレカルチャー
(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
(3)psB3K3(J04450)のトランスフォーメーション
【実験内容】
(1)プレカルチャー
:DH5α-pSB6を液体LB培地(+amp)2mLで培養
:DH5α-pSB1 を液体LB培地(+amp)2mLで培養
(2)pGVBpttの電気泳動
:〈組成〉
::pGVBppt 5μL
::KpmI 1μL
::HindⅢ 1μL
::Buffer(M) 1μL
::H2O 11μL
::計 20μL
:これを1h,37℃でインキュベート
: ↓
:Loading Buffer 1μLを加えたものと、マーカー(1kbp radder)を
:1% Agarose Gel のコームに流し込んだ
: ↓100V,30min泳動
: ↓EtBr(10mg/mL)で20min染色
:泳動結果を写真で撮影した
(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
: ↓
: 37℃でインキュベート（Overnight）
