(Difference between revisions)
|Line 46:||Line 46:|
* Macaya's and Bock's selection buffer: 20 mM Tris-acetate, pH 7.4, 140 mM NaCl
* Macaya's and Bock's selection buffer: 20 mM Tris-acetate, pH 7.4, 140 mM NaCl5 mM KCl1 mM CaCl<sub>2</sub>1 mM MgCl<sub>2</sub>
Revision as of 13:48, 10 July 2006
- Our goal is to to design and implement molecular containers, which can be dynamically opened and closed by an external stimulus.
- The containers will be implemented as DNA nanostructures, which afford a significant degree of positional control and chemical versatility.
- As an initial proof-of-concept, we plan to use our DNA containers to demonstrate controllable activation ("delivery") of anti-thrombin aptamers.
- We expect that molecular containers could have several interesting scientific and clinical applications, such as
- Drug and gene delivery
- Bio-marker scavenging (early detection of biomarkers)
- Directed evolution (compartmentalized selections)
- Using multiplexing for combinatorial chemical synthesis
- Capture and stabilization of multiprotein complexes
- Protein folding (chaperones)
- Cell sorting
Questions / procedures
- what percent gel? 10% to 20% polyacrylamide gels, no SDS (but would make for a good control)
- what incubation conditions?
- how much protein and DNA? protein at 1 μM, DNA at 2 μM
- Coomassie stain
- all combinations protein, aptamer, nanotube
- Macaya's and Bock's selection buffer: 20 mM Tris-acetate, pH 7.4, 140 mM NaCl, 5 mM KCl, 1 mM CaCl2, 1 mM MgCl2
- Liu's buffer: 40 mM Tris, 20 mM CH3COOH, 2mM EDTA, 12.5 mM (CH3COO)2Mg, pH 8.0
- Schultze P, Macaya RF, and Feigon J. . pmid:8107090.
- Liu Y, Lin C, Li H, and Yan H. . pmid:15945116.
- Li WX, Kaplan AV, Grant GW, Toole JJ, and Leung LL. . pmid:8298130.
- Bock LC, Griffin LC, Latham JA, Vermaas EH, and Toole JJ. . pmid:1741036.
- Macaya RF, Schultze P, Smith FW, Roe JA, and Feigon J. . pmid:8475124.