Takara ThruPLEX HV chemistry for DNA-seq and tag-seq from FFPE and cell-free DNA

ThruPLEX HV is a fast, modular NGS library preparation system engineered and optimized to generate DNA libraries with high molecular complexity and balanced GC representation from input volumes of 30 µl. This technology provides a complete, accurate system that enables reliable sequencing from challenging samples. The HV expansion of ThruPLEX technology accommodates a higher amount of starting material, with protocols now capable of handling inputs of 5 ng to 200 ng of dsDNA.

ThruPLEX HV is a fast, modular NGS library preparation system engineered and optimized to generate DNA libraries with high molecular complexity and balanced GC representation from input volumes of 30 µl. This technology provides a complete, accurate system that enables reliable sequencing from challenging samples. The HV expansion of ThruPLEX technology accommodates a higher amount of starting material, with protocols now capable of handling inputs of 5 ng to 200 ng of dsDNA. The entire three-step workflow takes place in a single tube or well, in about two hours, with minimal hands-on time. With high library diversity, ThruPLEX HV libraries excel when combined with target enrichment and deliver high-quality sequencing results. Our ThruPLEX DNA-Seq HV and ThruPLEX Tag-Seq HV kits demonstrate robust performance with FFPE and cfDNA and are offered with pre-plated, single-use UDI indexes for clear sample assignment.1, 2

The ThruPLEX HV PLUS kits are the newest members of the ThruPLEX HV family. The ThruPLEX HV PLUS Enzymatic Fragmentation Module is designed to perform size-tunable enzymatic fragmentation in tandem with the ThruPLEX HV repair step. The ThruPLEX DNA-Seq HV PLUS Kit and the ThruPLEX Tag-Seq HV PLUS Kit combine the ThruPLEX HV PLUS Enzymatic Fragmentation Module with the ThruPLEX HV Core Components. The ThruPLEX HV PLUS kits retain the remarkable features of the original ThruPLEX HV system and eliminate any need for mechanical or separate enzymatic fragmentation before ThruPLEX HV library prep.

Unlike other NGS library preparation kits based on ligation of Y-adapters, our patented ThruPLEX HV technology uses stem-loop adapters to construct high-quality libraries in a fast, efficient workflow. This protocol is the simplest in the industry and enables the addition of adapters containing Illumina®-compatible UDIs and UMIs in three short steps. The sample never leaves the tube, ensuring accurate sample tracking, minimizing handling errors, and preventing loss of valuable samples. Sensitive, even coverage is one of the hallmarks of this system, with both DNA-seq and tag-seq kits providing consistent, reproducible coverage for a broad range of GC content, without bias.

ThruPLEX Tag-Seq HV offers added power in correcting for amplification and sequencing errors—key advantages when detecting low-frequency variants. The UMIs included in the system have been carefully selected for even representation and the best possible performance both in error correction and sequencing results on Illumina platforms.

1The ThruPLEX HV UDI kits are not compatible with previous versions of ThruPLEX kits, including the ThruPLEX DNA-Seq, ThruPLEX Tag-Seq, and ThruPLEX Plasma-Seq kits. They are only compatible with the ThruPLEX HV or ThruPLEX HV PLUS kits.

2All indexes have been functionally validated to work with Illumina sequencing systems (e.g., MiSeq®, NovaSeqTM, MiniSeqTM, NextSeq®, and HiSeq® platforms) using two- or four-channel chemistry for base calling. They have not been validated with systems using one-channel chemistry.

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