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Team:HokkaidoU Japan/Notebook/August12
From 2010.igem.org
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* Checked the result of yesterdays ligation of PCR products by electrophoresis | * Checked the result of yesterdays ligation of PCR products by electrophoresis | ||
* Added 4 uL of 6x Sample Buffer to make 12 uL in total | * Added 4 uL of 6x Sample Buffer to make 12 uL in total | ||
| - | * Used | + | * Used 20 uL of EtOH |
<div style="clear:both"></div> | <div style="clear:both"></div> | ||
| - | =Mini prep= | + | =[[Team:HokkaidoU_Japan/Protocols|Mini prep]]= |
* Used cells incubated for 18h in LB broth | * Used cells incubated for 18h in LB broth | ||
| - | ** 1-18F didn't grow well | + | ** [[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 1-18F, but after centrifugation there were no precipitation | + | * 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 | ||
| - | =Electrophoresis | + | =Electrophoresis 2= |
[[Image:HokkaidoU Japan 20100812b.jpg|200px|right|thumb|Take 1]] | [[Image:HokkaidoU Japan 20100812b.jpg|200px|right|thumb|Take 1]] | ||
[[Image:HokkaidoU Japan 20100812c.jpg|200px|right|thumb|Take 2 after few minutes]] | [[Image:HokkaidoU Japan 20100812c.jpg|200px|right|thumb|Take 2 after few minutes]] | ||
| - | Checked plasmids gathered via mini prep and PCR products, by electrophoresis. <br> | + | Checked plasmids gathered via [[Team:HokkaidoU_Japan/Protocols|mini prep]] and [[Team:HokkaidoU_Japan/Protocols|PCR]] products, by electrophoresis. <br> |
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|style="border-right:1px solid #996;"| Parts | |style="border-right:1px solid #996;"| Parts | ||
| - | | 1-18F | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|1-18F]] |
| - | | 2-21H | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|2-21H]] |
| - | | 2-11P | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|2-11P]] |
| - | | 2-24G | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|2-24G]] |
| - | | 1-2M | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|1-2M]] |
| - | | 3-1E | + | | [[Team:HokkaidoU_Japan/Parts#BioBricks|3-1E]] |
|- | |- | ||
|style="border-right:1px solid #996;"| DNA solution | |style="border-right:1px solid #996;"| DNA solution | ||
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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] | * 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] | ||
* Marker in lane 1 ([https://2010.igem.org/Image:HokkaidoU_Pictures_DNA_Marker.png λ/''Hin''dIII]) leaked out | * 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 [[Team:HokkaidoU_Japan/Protocols|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 | ||
Latest revision as of 07:25, 27 October 2010
since 13 Jul 2010
since 1 Aug 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
