10X Genomics Chromium

The 10X Chromium library preparation system is a unique approach to leveraging single molecular barcoding. In the genomic DNA mode, the Chromium instrument uses molecular barcoding of high molecular weight DNA to generate long-read sequencing data using Illumina short read sequencing. In the single cell mode, the instrument can be used to generate single cell transcriptome data and profile an entire cell population.

Genomic DNA
Using high molecular weight DNA, (≥50kb) the Chromium instrument enables individual barcoding of long stretches of DNA. The long DNA fragments are then reduced to smaller sizes for Illumina short read sequencing, each retaining the barcode assigning it to the larger fragment. The libraries are constructed from these shorter fragments and sequencing on our Illumina HiSeq 2500s.

Applications

  • Long-range haplotyping, establishment of cis or trans relationships between variants
  • Generation of long range phase blocks
  • Detection of structural variants
  • Resolve genome sequencing for highly repetitive regions
  • De Novo assembly of novel diploid genomes
  • Exome Capture

Single Cell
Using the single cell mode on the Chromium instrument allows for the molecular profiling of a cell population. RNA transcripts from individual cells are barcoded to allow full transcriptome sequencing of individual cells. The instrument can profile from 1000 to 10,000 cells per experiment, which are then used to generate a single library for sequencing. Molecular barcodes assign individual transcript reads to a particular cell. Using the transcription profile, researchers can identify individual cell types in their population.

Applications

  • Interrogations of heterogeneous cell populations
  • Differential expression in cell populations
  • Determination of cell cycle phase

                                                    

Genomics

The David H. Murdock Research Institute (DHMRI) Genomics Laboratory features a range of services and platforms that assist our collaborators with all phases of their projects from DNA or RNA extraction to library preparation and next generation sequencing. Our Genomics Laboratory also provides microarray and quantitative PCR platforms. Our scientists’ expertise allows them to develop novel methods, assays and protocols that give our collaborators custom applications and technologies.  All of the work conducted in the Genomics Laboratory is supported by state-of-the art computational and bioinformatics resources that help our collaborators get the most from their genetic data.

Collaborators also benefit from the availability of complementary services provided by DHMRI laboratories that specialize in metabolomics, proteomics, NMR, microscopy and targeted biochemistry.

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