Next generation sequencing (NGS) has emerged as a powerful technique for elucidating the the diversity of the immune cell repertoire, but you can miss the longer clonotypes and incompletely characterize scFvs with short read approaches. The 150 nt or 300 nt Illumina reads are far from long enough to completely sequence a full-length T cell receptor, immunoglobulin, or scFv and, while long read platforms can provide the full-length sequence of a receptor's gene or transcript, they lack the accuracy needed for reliable, reproducible results.
Fortunately, Loop Genomics's synthetic long read technology is ideally suited for studying the immune cell repertoire. Loop Long-reads deliver highly accurate, unbiased, single molecule sequencing that can resolve and quantify subtly different clonotypes that other sequencing technologies miss. You can gain insight into which variable and constant regions an individual clonotype or scFv contains as a result of our 99.9959% sequencing accuracy and read lengths of up to 6 kb with our Amplicon Sequencing Service (3 kb in your own lab using the LoopSeq Amplicon kit and Illumina sequencers). And, unlike other sequencing kits and services, our technology is compatible with both DNA and RNA starting material.
Furthermore, you can profile immune cell transcriptomes and capture the full-length sequence of every transcript as well as obtain accurate transcript abundance measurements using our Single Cell Long-read Transcriptomics Service or LoopSeq Transcriptome Kit.
Through unparalleled sequence accuracy and bias-free single-molecule counting, Loop Long-reads dramatically change what you can learn about the adaptive immune system:
Understand the diversity of the immune cell repertoire with full-length, single cell sequencing of B cell receptors, T cell receptors, immunoglobulins, scFvs
Sequence genomic DNA or mRNA or both
Quantify dynamic changes to immune cell populations with bias-free single molecule counting
Characterize natural and synthetic immune repertoires easily and accurately
Find and follow rare clonotypes
Identify biomarker signatures
Answer questions that other technologies simply cannot address
Expand what you can discover about the adaptive immune response with Loop Long-reads.
Contact us if you have any questions about how our technology can advance your project or to make a service request.
How Loop Long-read sequencing works
Loop Long-read technology relies on the use of unique molecular identifiers (UMIs), aka barcodes, clever molecular biology, and efficient bioinformatics algorithms to assemble raw short read data from Illumina sequencers into single molecule reads of up to 6 kb in length.
The process looks like this:
ATTACH: Every sample is exposed to millions of unique barcodes, but only one barcode attaches per DNA molecule.
AMPLIFY: Every molecule, along with its unique barcode, is amplified using PCR.
DISTRIBUTE: The barcode is randomly distributed within the molecule.
SEQUENCE: Sequence the segment next to each barcode.
ASSEMBLE: Short reads that share the same barcode are combined algorithmically into a full-length molecule using linked-read de novo assembly.
Loop Long-read sequencing is a powerful way to enrich your understanding of the adaptive immune response:
Amplification happens after the UMI is added, quantitation of individual molecules is bias-free for accurate abundance measurements.
Because we assemble multiple reads for individual molecules, we can use consensus error correction resulting in the most accurate sequence of any platform on the market.