Fluid Operation
Binding DNA to Mica
146
MultiMode SPM Instruction Manual
Rev. B
1. Obtain the required materials:
•
Mica substrates
•
DNA: BlueScript II SK9(+) double stranded plasmid DNA, 2961 base pairs, 1mg/ml in
10mM Tris, 1mM ethylenediaminetetraacetic acid (EDTA) from Stratagene, La Jolla,
CA.
•
Buffer solution: 10 mM HEPES and 5 mM NiCl
2
pH 7.6 (for loose binding and air
imaging), or NiCl
2
(for tight binding and
fl
uid imaging)
2. Dilute DNA in buffer solution to a final concentration of 2.5 ng/µl.
3. Glue a piece of mica to a metal support as described in
mica substrate with a piece of adhesive tape. Place 30µl of the DNA solution in the center of
mica disk. The DNA will bind to the mica within 1 minute.
4. Load prepared sample onto the AFM scanner and assemble the fluid cell, as previously
. It may be helpful to wait for the temperature of the
buffer to stabilize (20 minutes or more) before imaging.
The sample is now ready for TappingMode imaging.
8.7.3 Acknowledgments
Veeco wishes to express its appreciation to the following individuals for sharing their experience to
assist in preparing this section: Monika Fritz, Manfred Radmacher, Magdalena Bezanilla, Helen G.
Hansma, Paul Hansma, Jason Cleveland, Jan H. Hoh, Serge Magonov, Chris Johnson, Don Hersch,
Tom Kovaleski, Gouliang Yang, Jamie Vesenka, and Eric Henderson.
