Anchor Binding Capacity

What you will need:

• Paramagnetic Streptavidin Beads
• Annealed Anchor

Binding Capacity: vary the amount of Anchor

• Make a series of 50 μL serial dilutions of 2 μM Anchor solution (1/2, 1/5, 1/10, 1/15, 1/20, 1/25, 1/30, etc)
• Prepare a tube for each dilution of Anchor made, each containing 40 μL 4 mg mL^-1 beads.
• Wash the beads twice with 100 μL Wash/Binding buffer and once with 100 μL water, then aspirate water off the beads.
• Add 20 μL of each dilution to it's respective tube with washed beads.
• Let bind at room temperature for 10 minutes.
• Take readings at A₂₆₀ with a UV-Vis Spectrophotometer of the dilutions prior to binding to bead and after binding to bead. (ng of DNA can also be found by in-gel band intensity quantification methods)
• Convert DNA measurements to pmol and plot pmol DNA versus mg of beads. The slope gives the binding capacity