Composite

Part:BBa_K2260002:Experience

Designed by: Rachelle Varga   Group: iGEM17_Calgary   (2017-10-25)
Revision as of 01:03, 2 November 2017 by Samwc98 (Talk | contribs) (→‎Applications of BBa_K2260002)


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Applications of BBa_K2260002

An assay was conducted in order to measure the effect of our part, Phasin-HlyA, on PHB secretion. Triplicates of E. coli BL21 containing either Phasin-HlyA and PhaCAB or only PhaCAB (as a control) were placed in LB media containing 3% glucose, and allowed to grow for 48 hours. PhaCAB, the gene that produces PHB, ensured that equal amounts of PHB were produced in both our test and the control. The PhaCAB part came from Imperial College 2013, and can be found at https://parts.igem.org/wiki/index.php?title=Part%3ABBa_K1149052. At 24 hour intervals, live cells were separated from the media using differential centrifugation. This resulted in two fractions: a cellular fraction containing the live cells, and a secreted fraction containing the PHB that was released into the media, either through secretion or cell death. Before separation, CaCl2 was added to agglomerate PHB particles together.

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UNIQff4e63e7d4c82aa4-partinfo-00000000-QINU UNIQff4e63e7d4c82aa4-partinfo-00000001-QINU