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Team:HokkaidoU Japan/Notebook/aggregation Week 4
From 2012.igem.org
(23th mini-prep 24th electrophoresis) |
(Ethanol precipitation) |
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<div class="hokkaidou-notebook-daily"> | <div class="hokkaidou-notebook-daily"> | ||
| - | ==July | + | ==July 23th== |
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==Mini-prep== | ==Mini-prep== | ||
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==Electrophoresis== | ==Electrophoresis== | ||
| + | <p> | ||
Electrophoresis for Ag43(mini-preped above) and Ag43 digestion results(digested with EcoRI and SpeI) | Electrophoresis for Ag43(mini-preped above) and Ag43 digestion results(digested with EcoRI and SpeI) | ||
[[image:|thumb|Mini-prep result]] | [[image:|thumb|Mini-prep result]] | ||
| - | [[image:|thumb|Digestion result]] | + | [[image:HokkaidoU2012 120724 Ag43(K346007 cut with E&S) digestion product.jpg|thumb|Digestion result]] |
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| + | In this digestion result, we knew that one or two enzymes didn't work successfully but there are enough concentration of DNA in 3000bp band to use in digestion. | ||
| + | |||
| + | </p> | ||
| + | ==Gel extraction== | ||
| + | <p> | ||
| + | Gel extraction for digestion production. We used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50ul of DNA solution. | ||
| + | </p> | ||
| + | ==Ethanol precipitation== | ||
| + | <p> | ||
| + | Ethanol precipitation for digestion and gel extraction product. | ||
| + | #Added 5ul of NaoAc, 1.5ul of glycogen and 125ul of 100% ethanol. | ||
| + | #Centrifuged in 15000rpm, 10min at 4C. | ||
| + | #Remove supernatant and added 220ul of 70% ethanol. | ||
| + | #Centrifuged in 15000rpm, 10min at 4C. | ||
| + | #Remove supernatant and air drying in room temperature then added 10ul of DW. | ||
| + | </p> | ||
| + | [[image:|thumb|Ethanol precipitation result]] | ||
</div> | </div> | ||
<!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> | <!-- DO NOT EDIT UNDER THIS LINE @iTakeshi --> | ||
Revision as of 06:56, 24 July 2012
Contents |
July 23th
Mini-prep
mini-prep for Ag43(7/17 cultivated colony resuspended and 7/20 cultivated colony resuspended). We used FastGene Plasmid Mini Kit(Nippon Genetics) and got 50ul of DNA solutions.
Electrophoresis
Electrophoresis for Ag43(mini-preped above) and Ag43 digestion results(digested with EcoRI and SpeI) [[image:|thumb|Mini-prep result]]
_digestion_product.jpg)
In this digestion result, we knew that one or two enzymes didn't work successfully but there are enough concentration of DNA in 3000bp band to use in digestion.
Gel extraction
Gel extraction for digestion production. We used FastGene Gel&PCR extraction kit(NipponGenetics)and got 50ul of DNA solution.
Ethanol precipitation
Ethanol precipitation for digestion and gel extraction product. #Added 5ul of NaoAc, 1.5ul of glycogen and 125ul of 100% ethanol. #Centrifuged in 15000rpm, 10min at 4C. #Remove supernatant and added 220ul of 70% ethanol. #Centrifuged in 15000rpm, 10min at 4C. #Remove supernatant and air drying in room temperature then added 10ul of DW.
[[image:|thumb|Ethanol precipitation result]]
