title: A simple filtration technique for obtaining purified human chromosomes in suspension.
abstract: Here we present a simple method for cleaning polyamine human mitotic chromosomes in solution. This was achieved by filtering intact (unburst) nuclei along with both large and small cytoplasmic debris through a series of different pore sized filters. Pure human chromosomes were recovered using a simple reverse filtration step. Fluorescence microscopy was used to validate the chromosome suspension after each filtration step. This reverse filtration technique is an improvement in both procedure time and chromosome recovery compared to currently used post-purification methods. Chromosomes purified by our method could be used for many applications, such as structural studies using microfluidics and high resolution imaging or generation of chromosome paints and sequencing after flow cytometry.
authors.affiliation: London Centre for Nanotechnology, University CollegeLondon, London, UK; Research Complex at Harwell (RCaH), Rutherford Appleton Laboratory, Oxon, UK.
authors.forename: Mohammed
authors.initials: M
authors.lastname: Yusuf
authors.affiliation: London Centre for Nanotechnology, University CollegeLondon, London, UK; Research Complex at Harwell (RCaH), Rutherford Appleton Laboratory, Oxon, UK.
authors.forename: Neha
authors.initials: N
authors.lastname: Parmar
authors.affiliation: London Centre for Nanotechnology, University CollegeLondon, London, UK; Research Complex at Harwell (RCaH), Rutherford Appleton Laboratory, Oxon, UK.
authors.forename: Gurdeep K
authors.initials: GK
authors.lastname: Bhella
authors.affiliation: London Centre for Nanotechnology, University CollegeLondon, London, UK; Research Complex at Harwell (RCaH), Rutherford Appleton Laboratory, Oxon, UK.
authors.forename: Ian K
authors.initials: IK
authors.lastname: Robinson
mesh_terms: D002877:Chromosomes, Human; D005374:Filtration / Q000379:methods*; D005821:Genetic Techniques; D006801:Humans; D008856:Microscopy, Fluorescence
keywords: chromosome; cytoplasmic debris; filtration; mitotic; nuclei