Team:HokkaidoU Japan/Notebook/August12

From 2010.igem.org

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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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* Used markers are [https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''dIII] and [https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png pUC119/''Hin''fI]
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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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* Marker in lane 1 ([https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''dIII]) leaked out
* Mini prep sample，1-18F band in lane 3 wasn't visible so we decided PCR it tomorrow
* 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
* In lane 4 there were some visible plasmid dimers and trimer s

Revision as of 06:55, 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 λ/HindIII and pUC119/HinfI
  • Marker in lane 1 (λ/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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