Team:HokkaidoU Japan/Notebook/September7

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==前日のligationに使用したDNAの濃度測定==
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* RFPはH bufferを使ったものもM bufferを使ったものも4 ng/uLくらい
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* pSB1C3はそれよりずっと薄くてわずかにしか見えない
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==エタ沈==
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= Checked concentration of DNA used yesterday for ligation =
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Mighty Mixの標準プロトコルによると，ゲル抽後のDNAはTEに溶かしてある方が良いらしいので，濃縮の意味も込めてエタ沈してから，Ligationしてみることにした
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* 4 ng/uL RFP digested using either H buffer and M buffer
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* pSB1C3 band was very week
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==コンピテントセルのPCR==
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= Ethanol precipitation =
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先日のコロニーPCRで謎なバンドがすべてにみられたので，大腸菌ゲノム由来のものである可能性をみるため，コンピテントセルをPCRにかけたところ，バンドはみられなかった
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* すべてにみられたバンドはベクター由来か
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In accordance to Mighty Mix default protocol that for ligation DNA must melted in TE and for concentrating it more Ethanol precipitation was done.
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= Colony PCR of competent cells =
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Previous colony PCR produced strange bands so checked if there were from DNA of competent cell
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* Colony PCR didn't produce any bands so those bands had to be an artifact from plasmid used for transformation

Latest revision as of 11:49, 27 October 2010

Notebook

Checked concentration of DNA used yesterday for ligation

  • 4 ng/uL RFP digested using either H buffer and M buffer
  • pSB1C3 band was very week

Ethanol precipitation

In accordance to Mighty Mix default protocol that for ligation DNA must melted in TE and for concentrating it more Ethanol precipitation was done.

Colony PCR of competent cells

Previous colony PCR produced strange bands so checked if there were from DNA of competent cell

  • Colony PCR didn't produce any bands so those bands had to be an artifact from plasmid used for transformation
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