Author: Hieu Bui Advisors: Bill Knowlton and Bernie Yurke 7/30 ...

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Author: Hieu Bui
Advisors: Bill Knowlton and Bernie Yurke
7/30/2009
Version 1.0
Protocol for using Colloidal Gold Conjugates Streptavidin with DNA origami
nanostructures
General safety procedures:
Colloidal gold conjugates should be stored with desiccant in a freezer at 2-6 C.
Vials containing colloidal gold conjugates should be protected from light, for
example, by wrapping with aluminum foil.
Preparation of the stock solution should be carried out as close to the date of use
as possible.
When handling DNA solutions, gold conjugates, and TAE buffer, use of personal
protective equipment is mandatory – Nitrile gloves, splash goggles, and lab coat
are kept in ET 109
Any sample prep that takes place in ET 109 will be done on the designated
sample prep area
After handling chemicals, washing hands is mandatory before eating or leaving
the lab area. When the work is done for the day and the chemicals are put away,
and the gloves and goggles are removed, wash hands again before going on to
other tasks
Copies of the MSDS for gold conjugates must be kept in both ET109 and
whatever lab is used for refrigerated storage and mixing solutions
Contaminated solid waste, such as used gloves, pipette tips, and centrifuge tubes
will be disposed of in the “Solid Waste” bucket and the lid will be replaced on the
bucket after waste is added. If the bucket is full, contact Environment Health and
Safety, Wendy Campbell @ 63303, to come pick it up.
Prepare 0.1 x solution from colloidal gold conjugates streptavidine stock solution
(10-fold dilution)
Supplies:
1. Protective gloves, splash goggles, and lab coat
2. Aluminum foil
3. 0.5 mL micro-centrifuge tubes
4. 5 – 50 µL pipette and pipette tips
5. 1X TAE buffer Mg++
6. Colloidal gold conjugates stock solution
Procedure:
1. Pipette 10 µL of colloidal gold conjugates stock solution into 0.5 mL microcentrifuge
tube
2. Pipette 90 µL of 1X TAE buffer Mg++ into (1)
3. Mix the sample throughout and label properly (i.e. Col-Au-10-folds)
4. Dispose pipette tips and used gloves in the Contaminated Solid Waste bucket
Functionalize colloidal gold conjugates with customized ssDNA sequence

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Author: Hieu Bui
Advisors: Bill Knowlton and Bernie Yurke
7/30/2009
Version 1.0
Supplies:
1. Protective gloves, splash goggles, and lab coat
2. Aluminum foil
3. 0.5 mL micro-centrifuge tubes
4. 5 – 50 µL pipette and pipette tips
5. Col-Au-10-folds solution
6. Customized ssDNA sequence
Procedure:
1. Pipette 20 µL of Col-AuNPs-10-folds solution into 0.5 mL micro-centrifuge tube
2. Pipette 20 µL of customized ssDNA sequence into (1)
3. Mix the solution throughout and label properly (i.e. Functionalized Au)
4. The sample is ready after overnight incubation
5. Dispose pipette tips and used gloves in the Contaminated Solid Waste bucket