User:Klare Lazor/Notebook/Chem-496-001/2011/10/05

From OpenWetWare

Jump to: navigation, search
Biomaterials Design Lab Main project page
Previous entry      Next entry


The purpose is to use different concentrations of Ru(bpy) to determine the intensity and where change in intensity stops being linear. In hopes to test the sensitivity of the fluorimeter.


Miniprep/ Harvesting DNA

Using the cell culture previously grown, the liquid from the cell culture was dumped and the cells were fully resuspended using 250µL of Cell Resuspension Solution and 250µL of Cell Lysis Solution. This new mixture of cells and solution was transferred to a centrifuge tube. 10µL of Alkaline Protease solution was added to the mixture and the tube was inverted 4 times to mix contents. The mix was incubated at room temperature for 5 minutes. 350µL of Neutralization solution was added, and the tube was inverted 4 times to mix the contents. The tube was centrifuged at top speed for 10 minutes. Liquid (NOT the solid gathered at the bottom due to centrifuging) in the tube was pipetted into a "spin column" and was vacuumed through the spin column using a Vaccum Adapter and a Manifold Port (specialized instrumentation). 750µL of Wash Solution (ethanol added) was used to wash the contents of the spin column by applying vacuum. This was repeated using 250µL of the Was Solution. The contents of the spin column were dried by vacuuming for 10 minutes. The column was placed inside a 2 mL collection tube, filter-side down, and was centrifuged at top speed for 2 minutes. The column was transfered to a sterile 1.5mL centrifuge tube. 100mL of Nuclease-Free Water was added to the column, and the column was centrifuged at top speed for 1 minute. The column was discarded and the contents of the centrifuge tube were labelled and stored at -20 degrees Celsius.


  • Add data and results here...


This area is for any observations or conclusions that you would like to note.

Use categories like tags. Change the "Course" category to the one corresponding to your course. The "Miscellaneous" tag can be used for particular experiments, as instructed by your professor. Please be sure to change or delete this tag as required so that the categories remain well organized.

Personal tools