Single Molecule Fluorescence Microscopy Measurement of BM21_172/173 with/without Spermine

Aim Seeing tendencies in changes in length of the origami when the intercalator spermine is added.

Preparation

• Preparation was done according to Immobilizing structures with biotin oligos for TIRF
• The spermine concentrations were set to
  • One molecule spermine per 7 bases (= 0.802 μM)
  • One molecule spermine per 21 bases (= 0.241 μM)
  • No spermine = control

For calculations see Calculation of intercalator concentrations.

Measurement

• The excitation time was set to 0.1s/frame.
• 500 Pictures per video were taken.

Results

The amount of FRET events found in Matlab analysis was too low for a reliable interpretation.

Observed TEM Grids

• 2B3 2.27µM EtBr: good stain and dilution