A Simple Filtration Technique for Obtaining Purified Human Chromosomes in Suspension
Author(s) -
Mohammed Yusuf,
Neha Parmar,
Gurdeep K. Bhella,
Ian Robinson
Publication year - 2014
Publication title -
biotechniques
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.617
H-Index - 131
eISSN - 1940-9818
pISSN - 0736-6205
DOI - 10.2144/000114168
Subject(s) - filtration (mathematics) , chromosome , microfluidics , chromatography , fluorescence microscope , suspension (topology) , biology , flow cytometry , fluorescence , chemistry , microbiology and biotechnology , materials science , genetics , nanotechnology , gene , optics , physics , mathematics , statistics , homotopy , pure mathematics
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.
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