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After several years of rapid development, the new generation sequencer finally slowed down. Nowadays, sample preparation has become a new bright spot in NGS. New reagents and instruments are constantly emerging to shorten the time and improve the flux. At the same time, new methods are constantly being developed to handle fewer samples, such as single cells. This time, Biolink takes you to NGS sample preparation to see what new tools can help us meet the challenge of sample preparation.
Even with various kits, NGS sample preparation is still a painful process, which is time-consuming and laborious. For a high-throughput laboratory, that kind of hard work, hey, you know. As a result, people have turned their attention to those high-end atmospheric grade automatic preparation platforms. These instruments can not only save us from tedious manual operations, but also reduce human errors. Therefore, although the price is high, it is worthwhile.
at present, many companies in the market have launched automated sample preparation platforms, including Beckman Coulter and PerkinElmer(Caliper). Of course, sequencer manufacturers are not far behind, and have launched their own supporting platforms. In February this year, Illumina released the key-press NeoPrep library preparation system at the AGBT conference.
Beckman Coulter
Beckman Coulter's NGS library preparation solutions include Spriworks Fragment Library System I-III, which are suitable for Illumina Genome Analyzer, Roche GS FLX and Life Tech SOLiD systems respectively. In addition, it also introduced Qualcomm's SPRIworks HT system, which is suitable for Illumina's NGS platform.
SPRIworks System I-III is a patented solid-phase reversible immobilization (SPRI) technology, which can complete the library preparation of up to 1 samples in 4-5 hours. This system consists of three parts: SPRI-TE nucleic acid extractor, method card for controlling liquid treatment, and cartridge containing all reagents needed for preparing the library. All reagents, tips and tubes required for the experiment are provided, so you only need to prepare samples and connectors. SPRIworks system automatically completes four key steps of sample preparation: terminal repair, adding A tail, joint connection, size selection and purification, without column purification and gel electrophoresis.
the p>SPRIworks HT system is based on Beckman Kurt's Biomek? FXP laboratory automation workstation. It not only has the same function as SPRIworks I-III, but also can realize the establishment, purification, standardization and sample merging of PCR system. Of course, with various optional accessories, you can also realize DNA shearing and PCR. In less than 6 hours, it can process 96 samples, and it only takes 3 hours if the size selection is eliminated. There are also several options for size selection: 15-35 bp, 25-45 bp and 35-7 bp. SPRIworks HT adopts a graphical user interface, which is quite simple to use.
Perkin Elmer (Caliper)
Caliper has always been the leader of Lab-on-a-Chip technology, and has rich experience in the field of automation. In recent years, it has also launched a number of innovative products in the upstream and downstream of NGS. In September 211, Caliper was acquired by PerkinElmer, and they joined forces to launch NGS Express last year. Workstation.
this is a small, flexible and easy-to-use library preparation system, which is suitable for desktop sequencers such as Ion Torrent PGM? And Illumina's MiSeq system. This product comes with a complete set of NGS sample preparation application packages that have been tested in practice. Therefore, even novice users can easily obtain a stable and high-quality library for sequencing, thus ensuring the quality of sequencing data. Of course, experienced users can also modify preset applications to meet individual needs.
The application of p>NGS Express is equipped with a graphical operation interface and a step-by-step setup wizard, which shows users the most intuitive guidance by combining pictures and texts from selecting the application, confirming the number of samples and placing the experimental products. The flux of the system is flexible, and each batch can handle 1-24 samples, which ensures the on-demand inspection and the shortest experimental period.
at the same time, PerkinElmer also developed sci clonngs workstation, which is suitable for high-throughput sequencing platforms such as HiSeq. This instrument can realize library preparation, sequence capture and standardization, and is suitable for mRNA-Seq, ChIP-Seq and other applications. Each run can process 8-96 samples, so that 48 libraries can be prepared every week, or 192 exons can be captured. In addition, PerkinElmer also provides the most cost-effective Zephyr NGS workstation, Post-PCR workstation, LabChip DS and LabChip GX for quantitative and quality control of nucleic acids.
Illumina
At this year's AGBT conference, Illumina also released a NeoPrep library preparation system. According to Illumina website, this instrument adopts digital microfluidic technology to realize key-press library preparation. It can obviously simplify the preparation process of TruSeq and Nextera libraries. Fewer steps, fewer opportunities for mistakes, which means more repeatability.
NeoPrep can also be integrated with Illumina's genome computing environment BaseSpace, providing a seamless process from sample preparation to sequencing to analysis. NeoPrep can bring 16 libraries ready for sequencing in each run, and some samples are as low as 1 ng. The first batch of NeoPrep kits will be TruSeq PCR-free and TruSeq Nano, followed by other TruSeq and Nextera kits. This instrument is expected to go on sale this summer.
of course, new products are still on the market. At the AGBT conference, GenCell Biosystems also introduced a fully integrated and automated desktop library preparation system CLiC LP? . According to reports, this instrument adopts GenCell's Composite Liquid Cell(CLC) technology, and simulates the natural cell structure by sealing all the reactions in oil, thus saving plastic consumables and saving time and cost. In the near future, I believe we will have more and better choices.