Editing BioMicroCenter:Research (section)

===Copy number analysis in single cells - [http://web.mit.edu/biology/www/facultyareas/facresearch/amon.html AMON LAB]- [http://web.mit.edu/biology Biology] and [http://web.mit.edu/ki KI]===
Accurate segregation of chromosomes is critical for maintenance of genomic stability.   Failure to properly segregate chromosomes results in an unbalanced chromosome number, a condition known as aneuploidy.  Aneuploidy for chromosome 21 is the cause of Down Syndrome, and the majority of solid tumors harbor aneuploidies for one or more chromosomes.  Apart from these diseases, it has also been reported that normal, untransformed cells missegregate chromosomes as often as 20% of divisions (Carere et al., 1999; Shi & King, 2005).  Furthermore, multiple groups have reported even higher levels of aneuploidy in the normal mammalian brain, with over 20% of neurons and glia being aneuploidy (Faggioli, Wang, Vijg, & Montagna, 2012; Pack et al., 2005; Rehen et al., 2001; 2005).  Traditionally, these phenomena have been studied using spectral karyotyping (SKY) and fluorescence in situ hybridization (FISH).  Unfortunately, these methods are susceptible to spreading and hybridization artifacts, respectively, and FISH is limited in the number of chromosomes that can be surveyed in a single experiment. The Amon laboratory has thus turned to single cell sequencing to provide a more robust and thorough assessment of aneuploidy in the brain and other tissues. <BR><BR>
Using tissue samples obtained from mice and humans, we isolated single cells by microaspiration and amplified genomic DNA using a commercially available kit.  We then relied on the BioMicro Center to prepare barcoded libraries and sequence these libraries using the Illumina HiSeq2000.  From sequencing reads, we were able to infer the copy number of each chromosome in a given cell.<BR><BR>