Highlights
- Fastest: Only 1.5 hours of hands-on time. No Tapestation® analysis or AMPure® clean-ups.
- Accurate:Â Utilization of real-time PCR limits PCR chimera formation.
- Increased Coverage:Â Novel primers increase phylogenetic coverage of Bacteria and Archaea and enable species-level resolution for human microbiome profiling.
Description
16S rRNA sequencing is a routine technique for microbiome composition profiling. Compared to shotgun metagenomics sequencing, 16S rRNA sequencing is more cost-effective and more robust; it generally requires less input DNA and is less impacted by the presence of host DNA. However, 16S rRNA sequencing has its own challenges. One major challenge is the formation of PCR chimeric sequences, which are artificial sequences resulting from the recombination of two or more PCR templates. Additionally, with common 16S primers, it is difficult to achieve both species-level resolution and broad phylogenetic coverage. Moreover, common 16S library preparation protocols used in the field have not been optimized to be cost-effective for large-scale applications. The D641 Quick-16S NGS Library Prep Kit aims to standardize the library preparation process for 16S rRNA sequencing. Distinguishing features of the kit are described below. Fastest 16S rRNA Library Prep. The D641 Quick-16S NGS Library Prep Kit utilizes real-time (quantitative) PCR (qPCR) rather than endpoint PCR for 16S rRNA amplification, enabling direct quantification of PCR products and eliminating the need for additional library quantification analysis such as TapeStation analysis or gel electrophoresis.
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