Difference between revisions of "Part:BBa K1807000:Experience"
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Above is a photograph of the BBa_K1807000 Gibson Assembly Reaction transformation. | Above is a photograph of the BBa_K1807000 Gibson Assembly Reaction transformation. | ||
Escherichia coli NEB 5 alpha cells were spread onto LB Agar Plates containing Chloramphenicol 34ng/mL, IPTG ng/mL, Xgal ng/mL. | Escherichia coli NEB 5 alpha cells were spread onto LB Agar Plates containing Chloramphenicol 34ng/mL, IPTG ng/mL, Xgal ng/mL. | ||
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| + | For the Gibson Assembly Reaction we mixed 100ng of PCR-linearized (provided with the distribution) and EcoRI-digested pSB1C3 vector backbone together with 3 times the molar ratio of our Adapter BioBrick G Block dsDNA. We incubated the reaction at 50 degrees Celsius for an hour and then used 2uL (out of 20uL total) in a standard heat-shock transformation of chemically competent cells. | ||
Revision as of 14:56, 18 September 2015
Above is a photograph of the BBa_K1807000 Gibson Assembly Reaction transformation. Escherichia coli NEB 5 alpha cells were spread onto LB Agar Plates containing Chloramphenicol 34ng/mL, IPTG ng/mL, Xgal ng/mL.
For the Gibson Assembly Reaction we mixed 100ng of PCR-linearized (provided with the distribution) and EcoRI-digested pSB1C3 vector backbone together with 3 times the molar ratio of our Adapter BioBrick G Block dsDNA. We incubated the reaction at 50 degrees Celsius for an hour and then used 2uL (out of 20uL total) in a standard heat-shock transformation of chemically competent cells.