Team:HokkaidoU Japan/Notebook/August12

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{{Template:HokkaidoU_Japan}}<div class="linkbar"><div class="prev">[[Team:HokkaidoU_Japan/Notebook/August11|August 11]]</div>[[Team:HokkaidoU_Japan/Notebook|Notebook]]<div class="next">[[Team:HokkaidoU_Japan/Notebook/August13|August 13]]</div></div>
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=Electrophoresis 1=
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[[Image:HokkaidoU Japan 20100812a.jpg‎|200px|right|thumb|Electrophoresis of yesterdays ligation of PCR products]]
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* Checked the result of yesterdays ligation of PCR products by electrophoresis
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* Added 4 uL of 6x Sample Buffer to make 12 uL in total
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* Used 20 uL of EtOH
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<div style="clear:both"></div>
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=[[Team:HokkaidoU_Japan/Protocols|Mini prep]]=
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* Used cells incubated for 18h in LB broth
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** [[Team:HokkaidoU_Japan/Parts#BioBricks|1-18F]] didn't grow well
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* Centrifuge after adding 430 uL of chloroform、collected supernatant
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* Added 2-propanol to [[Team:HokkaidoU_Japan/Parts#BioBricks|1-18F]], but after centrifugation there were no precipitation
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** Melted in 30 uL of TE and did electrophoresis
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=Electrophoresis 2=
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[[Image:HokkaidoU Japan 20100812b.jpg‎|200px|right|thumb|Take 1]]
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[[Image:HokkaidoU Japan 20100812c.jpg‎|200px|right|thumb|Take 2 after few minutes]]
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Checked plasmids gathered via [[Team:HokkaidoU_Japan/Protocols|mini prep]] and [[Team:HokkaidoU_Japan/Protocols|PCR]] products, by electrophoresis. <br>
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Because we wont do restriction enzyme digestion reaction system is ass follows.
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{| style="text-align:center" class="protocol"
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|-
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|style="border-bottom:1px solid #996; border-right:1px solid #996;"|'''Lane'''
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|style="border-bottom:1px solid #996;"| １
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|style="border-bottom:1px solid #996;"| ２
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|style="border-bottom:1px solid #996;"| ３
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|style="border-bottom:1px solid #996;"| ４
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|style="border-bottom:1px solid #996;"| ５
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|style="border-bottom:1px solid #996;"| ６
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|-
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|style="border-right:1px solid #996;"| Parts
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| [[Team:HokkaidoU_Japan/Parts#BioBricks|1-18F]]
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| [[Team:HokkaidoU_Japan/Parts#BioBricks|2-21H]]
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| [[Team:HokkaidoU_Japan/Parts#BioBricks|2-11P]]
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| [[Team:HokkaidoU_Japan/Parts#BioBricks|2-24G]]
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| [[Team:HokkaidoU_Japan/Parts#BioBricks|1-2M]]
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| [[Team:HokkaidoU_Japan/Parts#BioBricks|3-1E]]
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|-
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|style="border-right:1px solid #996;"| DNA solution
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| 2 uL
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| 2
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| 2
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| 1
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| 1
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| 1
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|-
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|style="border-right:1px solid #996;"| 6x Sample Buffer
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| 0.4 uL
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| 0.4
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| 0.4
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| 0.4
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| 0.4
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| 0.4
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|}
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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 λ/''Hin''dIII]) leaked out
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* Mini prep sample，1-18F band in lane 3 wasn't visible so we [[Team:HokkaidoU_Japan/Protocols|decided]] PCR it tomorrow
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* In lane 4 there were some visible plasmid dimers and trimer s

Latest revision as of 07:25, 27 October 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 20 uL of EtOH

Mini prep

  • Used cells incubated for 18h in LB broth
  • 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
Retrieved from "http://2010.igem.org/Team:HokkaidoU_Japan/Notebook/August12"