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User:DavidC/11 September 2009
From 2009.igem.org
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| + | === Friday the 11th === | ||
| + | |||
| + | ==== Ligation ==== | ||
| + | |||
| + | Ligation pairs plasmid / insert : <br> | ||
| + | BBa_B0014/ BBa_P1002<br> | ||
| + | BBa_B0030 / BBa_R0040 <br> | ||
| + | BBa_B0014 / BBa_C0012 <br> | ||
| + | |||
| + | First report: <br> | ||
| + | Plasmid = 1µL <br> | ||
| + | Insert = 4µL <br> | ||
| + | |||
| + | Second report: <br> | ||
| + | Plasmid = 1µL <br> | ||
| + | Insert = 2µL <br> | ||
| + | |||
| + | For each samples add sterilized water to obtain a maximum volume equals to 8µL. <br> | ||
| + | |||
| + | Ligation mix (NEB): <br> | ||
| + | 6µL T4 ligase + 6µL T4 ligase buffer. <br> | ||
| + | Add 2µL / tube. <br> | ||
| + | Incubation 1h at RT. <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> | ||
Revision as of 03:08, 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
Friday the 11th
Ligation
Ligation pairs plasmid / insert :
BBa_B0014/ BBa_P1002
BBa_B0030 / BBa_R0040
BBa_B0014 / BBa_C0012
First report:
Plasmid = 1µL
Insert = 4µL
Second report:
Plasmid = 1µL
Insert = 2µL
For each samples add sterilized water to obtain a maximum volume equals to 8µL.
Ligation mix (NEB):
6µL T4 ligase + 6µL T4 ligase buffer.
Add 2µL / tube.
Incubation 1h at RT.
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).
