The introduction of DNA sequencing instruments capable of producing millions of DNA sequence reads in a single run has profoundly altered the landscape of genetics and cancer biology. Complex questions can now be answered at previously unthinkable speeds and a fraction of their former cost. At the Sequencing Facility, NCI researchers are provided access to the latest technologies, with consultation and Q&A services available throughout the design and execution of sequencing projects.
Short Read Sequencing Technology
Illumina Sequencing
- Illumina sequencing utilizes reversible terminator chemistry optimized to achieve high levels of cost-effectiveness and throughput
- Millions of reads produced per sample lane at 50 bp to 300 bp read lengths
- Support for the multiplexing of 96 bar-coded samples into a single lane
- Available resources include NovaSeq6000, NextSeq500, Hiseq4000, and MiSeq sequencer
Long-Read Sequencing Technology
PacBio Sequel II Sequencing
- Amplification-free sequencing via single-molecule real-time (SMRT) technology enables rapid identification of long nucleotide chains, with average yields of 100-120 Gb per SMRT Cell for Continuous Long Reads (CLR – long insert libraries > 20 kb) or 200-400 Gb per SMRT cell for highly accurate Circular Consensus Sequence (CCS – short insert libraries < 20 kb).
- Polymerase read lengths averaging greater than 120,000 bases per molecule, with maximum read lengths > 200,000 bases, facilitate genome assembly and mapping of repetitive regions with high fidelity circular consensus reads.
- The option of 10 to 30 hour run times depending on desired data yield and accuracy, as well as project type.
- Project flexibility with production protocols that include large and small genome sequencing, whole and targeted transcriptome sequencing (Iso-Seq), and amplicon sequencing, either single or multiplexed.
Responsive design
- 10X Genomics Chromium System, powered by GemCode Technology, provides a precisely engineered reagent delivery method that enables thousands of micro-reactions in parallel.
- Incorporation of identifying barcodes enables reads generated by short read sequencing mapped back to their original HMW DNA, single cell, or single nucleus of origin.
- Genome and exome sequencing using Linked-Reads resolves genic phasing, reveals structural variation and detects variants in complex regions of the genome.
- Chromium Single Cell Gene Expression Solution enables cell characterization and gene expression profiling of thousands of cells, while Single Cell Immune Profiling enables simultaneous analysis
- Chromium Single Cell ATAC (Assay for Transposase Accessible Chromatin) and Single Cell CNV Solutions reveal the heterogeneity and regulatory landscape of the genome, providing insights into cell variability and clonal evolution.
Responsive design
- Bionano’s non-sequencing-based genome mapping technology images and analyzes extremely long, high-molecular-weight DNA
- Facilitate identification of structural variants and creation of de novo genome assemblies
We currently employ the following sequencing platforms:
Short Read Sequencing Technology
Illumina Sequencing
Illumina sequencing utilizes reversible terminator chemistry optimized to achieve high levels of cost-effectiveness and throughput
Millions of reads produced per sample lane at 50 bp to 300 bp read lengths
Support for the multiplexing of 96 bar-coded samples into a single lane
Available resources include NovaSeq6000, NextSeq500, Hiseq4000, and MiSeq sequencer
Long-Reads/PacBio Sequencing
PacBio Sequel II Sequencing
Amplification-free sequencing via single-molecule real-time (SMRT) technology enables rapid identification of long nucleotide chains, with average yields of 100-120 Gb per SMRT Cell for Continuous Long Reads (CLR – long insert libraries > 20 kb) or 200-400 Gb per SMRT cell for highly accurate Circular Consensus Sequence (CCS – short insert libraries < 20 kb).
Polymerase read lengths averaging greater than 120,000 bases per molecule, with maximum read lengths > 200,000 bases, facilitate genome assembly and mapping of repetitive regions with high fidelity circular consensus reads.
The option of 10 to 30 hour run times depending on desired data yield and accuracy, as well as project type.
Project flexibility with production protocols that include large and small genome sequencing, whole and targeted transcriptome sequencing (Iso-Seq), and amplicon sequencing, either single or multiplexed.
10X Genomics Chromium System, powered by GemCode Technology, provides a precisely engineered reagent delivery method that enables thousands of micro-reactions in parallel.
Incorporation of identifying barcodes enables reads generated by short read sequencing mapped back to their original HMW DNA, single cell, or single nucleus of origin.
Genome and exome sequencing using Linked-Reads resolves genic phasing, reveals structural variation and detects variants in complex regions of the genome.
Chromium Single Cell Gene Expression Solution enables cell characterization and gene expression profiling of thousands of cells, while Single Cell Immune Profiling enables simultaneous analysis
Chromium Single Cell ATAC (Assay for Transposase Accessible Chromatin) and Single Cell CNV Solutions reveal the heterogeneity and regulatory landscape of the genome, providing insights into cell variability and clonal evolution.
Bionano’s non-sequencing-based genome mapping technology images and analyzes extremely long, high-molecular-weight DNA
Facilitate identification of structural variants and creation of de novo genome assemblies