European Human Genetics Conference
Takara Bio will attend the 52nd annual European Human Genetics Conference from June 15–18, 2019 in Gothenburg, Sweden. The ESHG annual meeting gathers scientists from around the globe to present the latest advances in human genetics. We look forward to discussing our tools for genetic analysis research, including our gold-standard SMART-Seq Stranded and PicoPLEX Gold chemistries for single-cell RNA- and DNA-seq, respectively. Please stop by and meet us at Booth 240. Our highlights will include:
Poster: "A SMARTer solution to stranded single-cell RNA-seq"
With the growing need for low-input and single-cell NGS library prep solutions, we see that researchers recognize the value in revealing transcriptome profiles from damaged cells as well as noncoding information from extremely low cell numbers (1–1,000). We have previously released several RNA-seq solutions based on oligo(d)T priming that push the limits of sensitivity and reproducibility from ultra-low inputs as well as single cells. Oligo(d)T priming is a very efficient way to capture the transcriptome; however, only the polyadenylated fraction can be captured. In addition, for oligo(dT)-primed cDNA synthesis to generate high-quality libraries, one needs to start with high-quality RNA, which excludes the use of this technology with samples damaged or degraded due to the nature of the processing (e.g., FFPE samples) or method of isolation. Additionally, these earlier single-cell kits do not preserve strand-of-origin information. The SMART-Seq Stranded Kit now allows the generation of sequencing-ready, stranded Illumina libraries directly from 1–1,000 sorted cells or an equivalent amount (10 pg–10 ng) of purified total RNA of any quality. This kit integrates an innovative technology, already incorporated in our SMARTer Stranded Total RNA Seq Kit v2 - Pico Input Mammalian, which enables removal of ribosomal cDNA following cDNA synthesis, as opposed to direct removal of corresponding rRNA molecules prior to reverse transcription. The SMART-Seq Stranded Kit protocol can be completed within seven hours, and a convenient pooling option for inputs between one to ten cells facilitates greater ease-of-use by minimizing the number of samples being handled.
Poster: "Generation of single cell NGS library with high reproducibility, and greatly improved coverage and fidelity for precision medicine"
Accurate, reproducible detection of mutations and copy number variations (CNV) from small amounts of DNA, including single cells, is key for genetic analysis of clinical samples to assist in identifying the best treatment regimen and molecular diagnoses of diseases such as cancer. To allow accurate detection of both single nucleotide variants (SNVs) and CNVs, we have released an enhanced version of our PicoPLEX chemistry that improve sequencing coverage, uniformity, and accuracy while increasing the resolution for CNV detection and retaining reproducibility. This enhanced chemistry—named SMARTer PicoPLEX Gold Single Cell DNA-Seq Kit (PicoPLEX Gold)—is a single-cell library-prep kit with a simple, four-step protocol to convert single cells into NGS libraries in under three hours with minimum hands-on-time. Libraries prepared from single GM12878 cells using the PicoPLEX Gold kit were sequenced on an Illumina NextSeq® platform to a depth of ~35 million read pairs (2 x 150 cycles), generating >50% genome coverage. This coverage represents a 2-fold improvement over the original PicoPLEX kit, along with a 4X reduction in duplication rates. The PicoPLEX Gold kit detected 3.5X more SNVs compared to Multiple Displacement Amplification (MDA) with the same number of reads. The PicoPLEX Gold kit also produced up to 50% lower allele drop-in (false-positive) rates than MDA. The increased coverage and low bias of the PicoPLEX Gold kit translated to extremely low allele-dropout rates (ADO), ~5X lower than MDA. Therefore, a single PicoPLEX Gold kit library enables reliable, high-resolution CNV analysis with shallow sequencing, and an accurate and reproducible SNV and CNV analysis with deeper sequencing.
Posters will be discussed in four groups, from 10:15–11:15 and 16:45–17:45 on Sunday and Monday. All posters will remain on display from Saturday to Monday.
Prize draw—see us at booth #240 for details.
We're looking forward to seeing you at the meeting. In the meantime, we invite you to reach out to our scientists with any questions or requests via the "talk to a scientist" link below.
A novel protocol that enables the detection of both SNVs and CNVs from 1-5 cells in a single, low-pass sequencing run that saves time and money.
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