Rensselaer/29 July 2008

7/29/08

Cynthia, Anne, Chas

We removed supercompetent E. Coli cells (strain #?) from the -80 freezer and transformed them using puc-MN and puc-EBI and puc-crtY plasmids for expression of carotenoids. Cells with the crt-M and crt-N and genes are expected to synthesize 30-carbon carotenoids and display yellow pigmets and those with crt-E, crt-B, and crt-I are expected to synthesize 40-carbon carotenoids and display red or pink pigments. After an hour of shaking at 37 degrees C, the cells were plated on LB Agar with ampicillin. A plate containing 200uL of transformed cells and a plate containing 50uL of transformed cells was made for each plasmid. 500mL of LB Agar was mixed and autoclaved to make plates with both ampicillin and chloramphenicol for pac-EBI/puc-crtY transformations.