Team:HokkaidoU Japan/Notebook/August12
From 2010.igem.org
(Difference between revisions)
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=[[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/ | + | ** [[Team:HokkaidoU_Japan/Parts#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/ | + | * Added 2-propanol to [[Team:HokkaidoU_Japan/Parts#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 | ||
- | | [[Team:HokkaidoU_Japan/ | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|1-18F]] |
- | | [[Team:HokkaidoU_Japan/ | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|2-21H]] |
- | | [[Team:HokkaidoU_Japan/ | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|2-11P]] |
- | | [[Team:HokkaidoU_Japan/ | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|2-24G]] |
- | | [[Team:HokkaidoU_Japan/ | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|1-2M]] |
- | | [[Team:HokkaidoU_Japan/ | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|3-1E]] |
|- | |- | ||
|style="border-right:1px solid #996;"| DNA solution | |style="border-right:1px solid #996;"| DNA solution |
Latest revision as of 07:25, 27 October 2010
Electrophoresis 1
- 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
- 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
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