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User:DavidC/22 September 2009
From 2009.igem.org
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| + | === Tuesday the 22th === | ||
| + | |||
| + | ==== Ligation ==== | ||
| + | |||
| + | Ligation schemes: plasmid / insert: <br> | ||
| + | |||
| + | BBa_B0014 / BBa_P1003 ;<br> | ||
| + | BBa_B0014 / BBa_C0012. <br> | ||
| + | |||
| + | First report: <br> | ||
| + | Plasmid = 2µL <br> | ||
| + | Insert = 3µL <br> | ||
| + | |||
| + | Second report: <br> | ||
| + | Plasmid = 1µL <br> | ||
| + | Insert = 2µL <br> | ||
| + | |||
| + | Third report: <br> | ||
| + | Plasmide = 1µL <br> | ||
| + | Insert = 4µL <br> | ||
| + | |||
| + | ==== TAKARA DNA ligation kit ==== | ||
| + | |||
| + | First and third report: <br> | ||
| + | Solution A (Buffer) = 25µL <br> | ||
| + | Solution B (Enzyme) = 5µL <br><br> | ||
| + | |||
| + | Second report: <br> | ||
| + | Solution A (Buffer) = 15µL <br> | ||
| + | Solution B (Enzyme) = 3µL <br><br> | ||
| + | |||
| + | Incubation 1h at 16°C. <br> | ||
| + | |||
| + | ==== Electroporation ==== | ||
| + | |||
| + | Electroporation cuvettes = 2mm ; inoculums of electrocompetent <i>E.coli</i> DH5alpha= 40µL; pulse = 2,5KVolt ; 1h of incubation. <br> | ||
| + | |||
| + | Spread 1mL of inoculums into a petri dish with LB + ampicillin (50mg/mL) (20/0,02mL). <br> | ||
Latest revision as of 03:34, 21 October 2009
Introduction Current therapies Gene therapies DVS Concept Tissue Vector Cell Vector Therapeutic plasmid Conclusion Implementation Tissue targeting Antitumor action Treatment modeling Conclusion
BioBricks Material & Method Bibliography
Contents |
Tuesday the 22th
Ligation
Ligation schemes: plasmid / insert:
BBa_B0014 / BBa_P1003 ;
BBa_B0014 / BBa_C0012.
First report:
Plasmid = 2µL
Insert = 3µL
Second report:
Plasmid = 1µL
Insert = 2µL
Third report:
Plasmide = 1µL
Insert = 4µL
TAKARA DNA ligation kit
First and third report:
Solution A (Buffer) = 25µL
Solution B (Enzyme) = 5µL
Second report:
Solution A (Buffer) = 15µL
Solution B (Enzyme) = 3µL
Incubation 1h at 16°C.
Electroporation
Electroporation cuvettes = 2mm ; inoculums of electrocompetent E.coli DH5alpha= 40µL; pulse = 2,5KVolt ; 1h of incubation.
Spread 1mL of inoculums into a petri dish with LB + ampicillin (50mg/mL) (20/0,02mL).
