tSMS

SeqLL Technical Explanation

SeqLL, Inc. specializes in true single molecule sequencing of RNA or DNA. As the sole service provider for Helicos True Single Molecule Sequencing technology, SeqLL offers researchers the highest quality RNA sequencing data available by capturing the greatest depth and breadth of findings.

The Helicos process does not require ligation, PCR amplification, or lengthy library preparation procedures and, thereby, avoids bias, loss of complexity, and under-representation of rare transcripts. This simplicity streamlines sample preparation, allowing SeqLL to provide a more consistent, accurate, and quantitative representation of the samples’ genomic content. With SeqLL True Single Molecule Sequencing, every molecule has an equal chance of making it to the flow cell surface for imaging. As a result, SeqLL technology represents the de-facto standard for detection and measurement of non-coding RNAs and rarely expressed transcripts.

Fold changes as low as <2-3 X can distinguish a disease associated physiological process from a normal one. However PCR amplification in standard library preparations can render these small changes impossible to track quantitatively. The additional noise, error, and documented loss of original transcriptome complexity cause small changes to fail statistical thresholds with technologies that employ protracted library preparation. SeqLL’s True Single Molecule Sequencing, ideal for transcriptome counting applications, captures all expressed transcripts and not just those represented by abundant or highly differentially-expressed RNAs. Our approach is especially critical for complex samples (ie - tissues) consisting of multiple cell varieties where expression and/or induction of any given transcript may be limited to a particular cell type.

SeqLL True Single Molecule Sequencing technology incorporates a level of flexibility rarely seen in sequencing platforms and allows researchers to innovate with special applications which will yield highly informative datasets. We offer direct sequencing of polyA+ RNAs from total RNA. After cells are lysed, without further steps, we anneal polyA+ and RNAs to the surface of the flow cell and sequence.

SeqLL’s read lengths range from 25 bases to over 60 with 35 being the average. While our reads are shorter, our sensitivity and accuracy is unparalleled. SeqLL can sequence 50 channels in a run and we can accept any number of samples. This allows us to avoid multiplexing, which can cause further degradation of the original signal, distorting the quality of the measurements, and resulting in reduced accuracy and reproducibility.

True Single Molecule Sequencing technology emphasizes high quality findings rather than merely large quantities of uneventful reads. Our results are highly correlated and reproducible with >0.99 R^2 values and are very accurate to 1 read per million or less. Precise, unbiased measurements and low sample quantities are the strengths of True Single Molecule Sequencing. We, at SeqLL, are very effective with sequencing the likes of native viral genomes and immune-precipitated methylated DNA. We sequence independent molecules and provide informative content rather than sequencing copies of copies. SeqLL’s native nucleic acids and quantitative approach ensures success with accurate detection of small changes in complex cell types.

SeqLL - Sequencing the Lower Limit – Sensitive, Accurate, Direct, Reliable – Single molecule sequencing technology is available to support your research needs.