Team:HokkaidoU Japan/Notebook/September8

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(Difference between revisions)
(コロニーPCR)
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=コロニーPCR=
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=Colony PCR=
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PS-R, EX-FによるコロPがうまくいくのかを確かめるため&目的のinsertionを確かめるためのコロP
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Checked if  primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids
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* 6日のトラフォメ H buffer×2,M buffer×2
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* September 6th transformation (digestion was done with H buffer×2 and M buffer×2, 2 version)
* 1-3A
* 1-3A
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* 6日にやったトラフォメをもう一度
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* Retried September 6th transformation
* pUC119
* pUC119

Revision as of 17:57, 27 September 2010

Colony PCR

Checked if primers PS-R, EX-F were good to use for colony PCR and also checked if insert was realy in plasmids

  • September 6th transformation (digestion was done with H buffer×2 and M buffer×2, 2 version)
  • 1-3A
  • Retried September 6th transformation
  • pUC119

pSB1C3の確認

1-3A(RFP reporter)由来のpSB1C3を使っているため,RFP発現がこれに由来している可能性が無くない

  • H buffer処理 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
  • M buffer処理 2 uL + Mighty Mix 2 uL + T4 ligase 0.5 uL → Ligation → Transformation
  • H buffer処理 2 uL → Transformation
  • H buffer処理 2 uL → Transformation

GFPのPCR

  • GFP reporter BBa_I13522 pSB1A2 2-8A 937bp
  • GFP protein BBa_E0840 pSB1A2 1-12O 878bp
9月1日と同じメニューでPCRを行った