The isolation of single types of chromosomes is a critical technology for a wide variety of genome related research projects. The challenge is to recover large numbers of chromosomes with high purity and high molecular weight in a relatively short period of time. Our approach has been to develop an optical sorter for high speed sorting and to adopt commercial sorters (EPICS V and EPICS 752) for conventional chromosome sorting. All three units have dual laser excitation for Chromomycin A3 and Hoechst 33258 bound to the chromosomes. The optical chromosome sorter/selector is capable of sorting rates approximately 30-50 times faster than commercial sorters. With the recent success of cloning very large pieces of DNA (>100 kb) into a variety of large insert cloning vectors, the need for high throughput has become crucial. We have validated the technique of optical sorting (selection) using two different methods of damaging DNA of unwanted chromosomes to a degree that it does not be clone; either by cross-linking or by fragmentation of the DNA to size fragments too small for cloning. Both techniques have been shown to be capable of generating chromosome specific cosmid libraries with greater than 95% purity.
| Frumkin, Jesse P; Patra, Biranchi N; Sevold, Anthony et al. (2016) The interplay between chromosome stability and cell cycle control explored through gene-gene interaction and computational simulation. Nucleic Acids Res 44:8073-85 |
| Johnson, Leah M; Gao, Lu; Shields IV, C Wyatt et al. (2013) Elastomeric microparticles for acoustic mediated bioseparations. J Nanobiotechnology 11:22 |
| Micheva-Viteva, Sofiya N; Shou, Yulin; Nowak-Lovato, Kristy L et al. (2013) c-KIT signaling is targeted by pathogenic Yersinia to suppress the host immune response. BMC Microbiol 13:249 |
| Ai, Ye; Sanders, Claire K; Marrone, Babetta L (2013) Separation of Escherichia coli bacteria from peripheral blood mononuclear cells using standing surface acoustic waves. Anal Chem 85:9126-34 |
| Sanders, Claire K; Mourant, Judith R (2013) Advantages of full spectrum flow cytometry. J Biomed Opt 18:037004 |
| Cushing, Kevin W; Piyasena, Menake E; Carroll, Nick J et al. (2013) Elastomeric negative acoustic contrast particles for affinity capture assays. Anal Chem 85:2208-15 |
| Piyasena, Menake E; Austin Suthanthiraraj, Pearlson P; Applegate Jr, Robert W et al. (2012) Multinode acoustic focusing for parallel flow cytometry. Anal Chem 84:1831-9 |
| Austin Suthanthiraraj, Pearlson P; Piyasena, Menake E; Woods, Travis A et al. (2012) One-dimensional acoustic standing waves in rectangular channels for flow cytometry. Methods 57:259-71 |
| Vuyisich, Momchilo; Sanders, Claire K; Graves, Steven W (2012) Binding and cell intoxication studies of anthrax lethal toxin. Mol Biol Rep 39:5897-903 |
| Chaudhary, Anu; Ganguly, Kumkum; Cabantous, Stephanie et al. (2012) The Brucella TIR-like protein TcpB interacts with the death domain of MyD88. Biochem Biophys Res Commun 417:299-304 |
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