Fractionation of chromosome 15 with an affinity-based approach using magnetic beads.
The two main shortcomings of the state-of-the-art method of sorting chromosomes, specificity and the efficiency of fractionating a significant amount of chromosomes, are addressed by this work in the design of a massively parallel approach using magnetic beads binding to a chromosome-specific DNA probe. In an attempt to isolate human chromosome 15 from a lymphoblastoid cell line, a chromosome 15 centromere-specific DNA probe with a fluorescent tag attached was reacted with the chromosomes. Magnetic beads bound to anti-FITC antibody were reacted with the labeled pool of chromosomes and separated by exposure to a magnetic field. The specificity of the fractionated pool was verified by performing fluorescence in situ hybridization on the isolated pool. The chromosome of interest could be enriched to about 75% within a maximum of 3-4 days, regardless of the amount of material.
Vitharana
Samadhi N
SN
Department of Chemistry, University of Kansas, 1251 Wescoe Hall Drive, Lawrence, KS 66045, USA.
Wilson
George S
GS
D045744:Cell Line, Tumor; D002503:Centromere / Q000737:chemistry* / Q000235:genetics; D002884:Chromosomes, Human, Pair 15 / Q000737:chemistry* / Q000235:genetics; D015342:DNA Probes / Q000737:chemistry* / Q000235:genetics; D006801:Humans; D017404:In Situ Hybridization, Fluorescence / Q000379:methods; D008863:Microspheres
D015342:DNA Probes