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Meeting, DNA/AuNP Attachment
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July 12, 2013
Notes from Meeting
- Adjust range and use SymphoTime to better fit FCS graphs in Igor
- Use error bars on calibration graphs (for Igor <N> values)
- Focus on attaching AuNPs to DNA & hybridizing MB to run on FCS
- Use a range of volumes of TES in MHA silica-coating procedure to change silica-coating thickness
- Take absorbance data to see differences between solution, then use TEM imaging to verify presence of silica-coating
DNA and AuNP Attachment Procedure
Calculations
- Done by Allison Alix.<br.>
- Extraction Procedure:
- Combine 250uL 4.45uM DNA (diluted in PBS) with 250uL 100nM DTT in 4% TEA
- React for 10 minutes
- Extract DTT with 4 2mL aliquots of ethyl acetate (DTT will partition into ethyl acetate, DNA will partition into aqueous solution)
- Combine 231.3uL 2.226uM DNA, 231.3uL 29.67nM 14.7nm AuNPs, and 37.3uL sodium citrate buffer prepared 07/11/2013 <br.>
- Final Concentrations in Solution:
- DNA: 1.03uM
- AuNP: 13.73nM
- Sodium citrate: 20mM
- React for 10 minutes
- Centrifuge 4 times, extracting the supernatant each time and then redispersing in 250uL 50nM HEPES buffer
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