Transfections

Flowcytometry

Detecting LovTAP

Talk about shifting peak (and why absent in PBS one: dsRed is degraded by formaldehyde)

Characterizing LovTAP

Non-leakage DsRed, where art thou?

Melanopsin Calcium Imaging

By using timelapse photography and fluorescence microscopy we were able to study the influx of calcium into CHO cells. The cells were stained with a calcium activated dye that fluoresces green, oregon green ( OGB-1/AM ), and exposed to blue light under the microscope.

The CHO seed cells show the expected response to light exposure. The dye is bleached and its intensity gradually decreases along an exponential curve over time. This would seem to indicate the concentration of calcium in the cell remained constant during light exposure and the dye was gradually bleached.

The CHO cells transfected with PHY42 contain human melanopsin. The cells show a slow linear decrease in dye intensity. This would seem to indicate that calcium is being imported into the cytoplasm while the dye is being bleached.

[http://partsregistry.org/wiki/index.php?title=Part:BBa_K838002]

Part assembly

We planned to clone several parts we used into pSB1C3, which is the standard backbone plasmid in which parts are shipped to the [http://partsregistry.org/Main_Page Registry]. Four parts were successfully inserted based on colony PCR results followed by successful sequencing.

LovTAP-VP16: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K838000 BBa_K838000]

Successfully cloned 4th of September.

LovTAP readout (RO): [http://partsregistry.org/wiki/index.php?title=Part:BBa_K838001 BBa_K838001]

Successfully cloned on 18th of September

Melanopsin: [http://partsregistry.org/wiki/index.php?title=Part:BBa_K838002 BBa_K838002] Successfully cloned on 12th of September