0 50 100 150 0 50 100 150 [4] KAPA Biosystems (2014) KAPA Library Quantification Technical Guide (KAPA Library Quantification Kits for Illumina® platforms) Find out more on our website: www.jnmedsys.com Need technical support? Pre-diluted DNA standards and freshly diluted library samples are prepared in triplicate and analyzed by qPCR. It was engineered specifically for use in SYBR Green-based qPCR through a process of directed evolution. Principle of the KAPA Library Quantification Assay. Please refer to the KAPA Library Quantification Technical Guide for more information. Results from Collibri and KAPA qPCR-based library quantification assays were also highly correlated (Figure 2). KAPA Library qPCR Quantification (product information) KAPA Library Quantification Kit technical data sheet; Illumina iScan. Kapa Library quantification kits for Illumina platforms. Product Compatibility Accurate quantification of Next Generation Sequencing (NGS) libraries is crucial for reliable clonal amplification via either emulsion or bridge PCR. Libraries are diluted to fall within the dynamic range of the KAPA Library Quantification assay (20 – 0.0002 pM). Next-generation sequencing library quantification; What is the enzyme in the KAPA SYBR ® FAST qPCR Master Mix (2X) Kits? Cell Prep Guide ; Sample and Library Preparation; 10x Chromium Long Read. qPCR primers are included and work with all Illumina adapters. Biomolecular Detection and Quantification, 2017, 14, 7-18. Standard qPCR library quantitation kits are available for all platforms from several vendors, and many researchers choose to prepare their own qPCR reagents. There are different methods for quantification and quality control depending on the sequencing library kit being used. We have demonstrated that the Collibri SS104001, A179, 4/16 4) Kapa Library quantification technical guide. KAPA SYBR DNA Polymerase is an engineered variant of Taq DNA polymerase. Please refer to the KAPA Library Quantification Technical Guide for more information. (qPCR). Contact our scientists: techsupport@jnmedsys.com For Research Use Only. Different qPCR kits are available for different qPCR chemistries, such as DNA binding dyes (eg, SYBR Green) and hydrolysis probes (eg, TaqMan probes). Product Compatibility The use of an automated liquid handling system is highly recommended for high-throughput NGS pipelines. Accurate quantification and proper quality check of next-generation sequencing libraries is key to a successful sequencing run. 3) Kapa Library quantification kits. List of assay types ; iScan Infinium Technology (video) 10x Chromium Single Cell. Next-generation DNA Sequencing meets Next-generation qPCR. Illumina recommends using KAPA® Library Quantification Kits for its sequencing systems. The KAPA Library Quantification assay consists of repetitive pipetting steps, which can easily be automated. The Illumina qPCR guide recommends using KAPA qPCR kits, which include six DNA standards to generate the standard curve. Technical Note: Understanding library quantification assays for next-generation sequencing applications NGS libraries must be carefully quantified and normalized so … across both library quantification kits (Figure 1). The KAPA Library Quantification assay consists of repetitive pipetting steps, which can easily be automated. Not for use in diagnostic procedures. Conclusions Library quantification is a crucial step in an NGS workflow, which helps ensure that each sample is sequenced to the desired depth. The use of an automated liquid handling system is highly recommended for high-throughput NGS pipelines.