User:Madeleine Y. Bee/Notebook/Single Molecule Fluorescence/2013/06/10

From OpenWetWare
< User:Madeleine Y. Bee‎ | Notebook‎ | Single Molecule Fluorescence‎ | 2013‎ | 06
Revision as of 08:43, 10 June 2013 by Madeleine Y. Bee (talk | contribs) (New: Modifying Silica-Coated AuNPs with HS-DNA Procedure)
Jump to: navigation, search
Owwnotebook icon.png Silica-coating Reaction <html><img src="/images/9/94/Report.png" border="0" /></html> Main project page
<html><img src="/images/c/c3/Resultset_previous.png" border="0" /></html>Previous entry<html>&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;</html>Next entry<html><img src="/images/5/5c/Resultset_next.png" border="0" /></html>

June 10, 2013

  • Silica-coating procedure from 05/24/2013 performed<br.>
    • MHA attachment performed, silica-coating reaction to be performed tomorrow

Buffer Preparation and Calculations

Sodium phosphate buffer, pH 9, to be used for the modification of HS-DNA and AuNPs@SiO2 procedure.<br.> monosodium phosphate- MW: 119.98g/mol<br.> disodium phosphate- MW: 141.96g/mol, pKA: 6.82<br.> buffer- pH 9,concentration 10mM<br.> <br.> 9=6.82+log([base]/[acid])<br.> 151.36=(10-[base])/([acid])<br.> [acid](151.36+1)=10<br.> [acid]=0.066mM<br.> [base]=9.935mM<br.> <br.> Use 0.0094g NaH2PO4 and 1.19188g Na2HPO4 in 1L water.<br.>

New: Modifying Silica-Coated AuNPs with HS-DNA Procedure


Chemicals Needed and Safety


  • Combine 150μL AuNPs@SiO2 (prepared 06/10/2013), 15μL APS and 235μL ethanol
  • Vortex solution for 30 minutes
  • Rinse sample three times by centrifugation at 7700g with ethanol and two times with sodium phosphate buffer, pH 9 (SPB)
  • Make a separate solution of 1 mg sulfo-SMCC in 400μL of SPB
  • Add SSMCC solution to AuNP solution
  • Vortex for 1 hour
  • Rinse twice with SPB
  • Rinse twice with potassium phosphate buffer, pH 7.4 (KPB)
  • Combine sample with 50μL 20μM HS-DNA
  • Vortex for 1 hour
  • Rinse three times with KPB
  • Re-suspend in 150μL KPB