Team:HokkaidoU Japan/Notebook/August12

From 2010.igem.org

(Difference between revisions)
Line 12: Line 12:
=[[Team:HokkaidoU_Japan/Protocols|Mini prep]]=
=[[Team:HokkaidoU_Japan/Protocols|Mini prep]]=
* Used cells incubated for 18h in LB broth  
* Used cells incubated for 18h in LB broth  
-
** [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-18F]] didn't grow well
+
** [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-18F]] didn't grow well
* Centrifuge after adding 430 uL of chloroform、collected supernatant
* Centrifuge after adding 430 uL of chloroform、collected supernatant
-
* Added 2-propanol to [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-18F]], but after centrifugation there were no precipitation
+
* Added 2-propanol to [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-18F]], but after centrifugation there were no precipitation
** Melted in 30 uL of TE and did electrophoresis
** Melted in 30 uL of TE and did electrophoresis
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|-
|-
|style="border-right:1px solid #996;"| Parts
|style="border-right:1px solid #996;"| Parts
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| [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-18F]]
+
| [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-18F]]
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| [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|2-21H]]
+
| [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|2-21H]]
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| [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|2-11P]]
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| [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|2-11P]]
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| [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|2-24G]]
+
| [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|2-24G]]
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| [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|1-2M]]
+
| [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|1-2M]]
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| [[Team:HokkaidoU_Japan/Material_And_Methods#Materials_And_Methods|3-1E]]
+
| [[Team:HokkaidoU_Japan/Materials_And_Methods#BioBricks|3-1E]]
|-
|-
|style="border-right:1px solid #996;"| DNA solution
|style="border-right:1px solid #996;"| DNA solution

Revision as of 11:26, 16 October 2010

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
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