User:Helen L. Slucher/Notebook/CHEM 571/2013/08/28: Difference between revisions

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==BSA-Au Nanoparticles==
==BSA-Au Nanoparticles==


Add 1mL of the (~2.5mM -note the exact concentration) gold (HAuCl4) solution to a 10mL volumetric flask
1. Add 1mL of the (~2.5mM -note the exact concentration) gold (HAuCl4) solution to a 10mL volumetric flask
Add an appropriate amount of BSA solution so that the final concentration of gold is 90X that of BSA
2. Add an appropriate amount of BSA solution so that the final concentration of gold is 90X that of BSA
Add deionized water up to 10mL
3. Add deionized water up to 10mL
Transfer solution to a test tube and cap with aluminum foil
4. Transfer solution to a test tube and cap with aluminum foil
Heat in oven at 80C for 3 hours
5. Heat in oven at 80C for 3 hours
Transfer solution to a plastic falcon tube (with blue cap)
6. Transfer solution to a plastic falcon tube (with blue cap)


Stock solutions made
Stock solutions made

Revision as of 15:15, 11 September 2013

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Entry title

Overview

Synthesize two different sets of gold nanoparticles. In one set, Au3+ is reduced by a protein (bovine serum albumin, BSA) and the synthesized nanoparticle is also surrounded and stabilized by BSA. In the second set, Au3+ is reduced by citrate, and the AuNP is stabilized by citrate in solution. The BSA-AuNPs are purple in color and the citrate-AuNPs are more of a burgundy (reddish) color.

BSA-Au Nanoparticles

1. Add 1mL of the (~2.5mM -note the exact concentration) gold (HAuCl4) solution to a 10mL volumetric flask 2. Add an appropriate amount of BSA solution so that the final concentration of gold is 90X that of BSA 3. Add deionized water up to 10mL 4. Transfer solution to a test tube and cap with aluminum foil 5. Heat in oven at 80C for 3 hours 6. Transfer solution to a plastic falcon tube (with blue cap)

Stock solutions made Gold solution (HAuCl4·3H2O) 0.0100g in 0.0100mL water → 2.54mM BSA solution 0.0104g BSA (MW = 66776g/mol) in 0.0100mL water → 15.6μM

Procedure

  1. In a PCR tube, mix the components on ice in the order they are listed above.
  2. Perform thermocycling program
    1. 95 °C 5 min
    2. 95 °C 30 s
    3. TH 30 s
    4. 72 °C 1 min for each 1 kb PCR product
    5. Repeat steps 2-4 a total of 12-36 times (24 is standard).
    6. 72 °C 5 min
    7. 12 °C hold

Notes

Please feel free to post comments, questions, or improvements to this protocol. Happy to have your input!

  1. List troubleshooting tips here.
  2. You can also link to FAQs/tips provided by other sources such as the manufacturer or other websites.
  3. Anecdotal observations that might be of use to others can also be posted here.

Please sign your name to your note by adding '''*~~~~''': to the beginning of your tip.

References

Relevant papers and books

  1. Goldbeter, A and Koshland, DE (1981) - Proc Natl Acad Sci U S A 78(11) 6840-4 PMID 6947258
  2. Jacob, F and Monod, J J (1961) - Mol Biol 3(3) 318-56 PMID 13718526
  3. Ptashne, M (2004) Genetic Switch: Phage Lambda Revisited - Cold Spring Harbor Laboratory Press ISBN 0879697164
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