User:Michael F. Nagle/Notebook/Chem 571/2012/10/17: Difference between revisions
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== | ==Electrophoresis== | ||
* | *5 μL of the plasmid mutated last week was placed in an autoclaved microcentrifuge tube. | ||
*1 μL 6x blue gel loading dye was added to this. | |||
*1 μL DPN1 was added to the other tube containing 45 μL plasmid solution. | |||
*This solution was placed on a VWR heat block at 37°C. | |||
*Agarose gel was prepared by adding 1.23g agarose to 35mL TAE buffer and heating via microwave for 35 seconds. | |||
*The agarose was poured into a Horizon 58 electrophoresis tray. | |||
*Once the gel had solidified after 30 minutes, the comb was removed. | |||
*The tray was filled with TAE buffer. | |||
*The plasmid and dye were added to wells. | |||
*Electrophoresis was run and the instrument was powered with a Bio-Rad Powerpac 200. | |||
*Upon completion of electrophoresis, 20μL 12.3g/mL ethidium bromide was added and slowly mixed with a VWR shaker. | |||
*The gel was viewed under UV light. | |||
==Chemiluminescence== | |||
*Because the fluorimeter was not working,[[User:Mary_Mendoza/Notebook/CHEM_571_Experimental_Biological_Chemistry_I/2012/10/17|Mary Mendoza]] tried to measure chemiluminescence of an HRP assay with luminol as prepared [[User:Michael_F._Nagle/Notebook/Chem_571/2012/10/16|yesterday]] using a UV/Vis instrument with the source lamp blocked, set to measuring energy with a slit width of 5nm. There was no signal. | |||
==Nucleation of AuNPs with BSA and Lysozyme== | |||
*[[User:Mary_Mendoza/Notebook/CHEM_571_Experimental_Biological_Chemistry_I/2012/10/17|Mary Mendoza]] prepared lysozyme and gold stock solutions. | |||
* Solutions for both BSA and lysozyme and HAuCl<sub>4</sub> were prepared at the same mole ratios that [[User:Michael_F._Nagle/Notebook/Chem_571/2012/08/29|have previously been used with BSA]]. Since the same BSA stock was used, and the lysozyme stock was prepared at the same concentration, the volumes added to each tube did not change. | |||
**Both sets of solutions were capped with foil and heated at 85<sup>o</sup>C for 4 hours. | |||
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Revision as of 01:17, 7 December 2012
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Electrophoresis
Chemiluminescence
Nucleation of AuNPs with BSA and Lysozyme
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