Description GenNext™ NGS Library Quantification Kit is for the SYBR® Green I qPCR-based library quantification of Illumina next-generation sequences. The kit allows the specific and accurate quantification of libraries bearing P5 and P7 adaptors which can be applied to flow cell amplification. It uses the highly efficient qPCR master mix KOD SYBR® qPCR Mix. Features • ACCURATE QUANTIFICATIONKOD SYBR® qPCR Mix can efficiently amplify GC- and AT-rich fragments of different lengths without bias. • BROAD DYNAMIC RANGEThe kit has a broad dynamic range from 20 pM (Standard DNA 1) to 0.0002 pM (Standard DNA 6). • CONVENIENtThe kit contains all reagents (KOD SYBR® qPCR Mix, 5x Primer Mix, Standard DNA, and 50x Dilution buffer) needed for the qPCR-based quantification of an NGS library. Application ・ NGS Library quantification for illumina’s insturment (ex. MiniSeq, MiSeq, HiSeq, NextSeq) Specification Category Item Details Storage condition - Store at -20℃ Components IKOD SYBR® qPCR mix* Details KOD SYBR® qPCR mix 1.67 mL x 3 50x ROX reference dye 250 µL Standard & Primer Set Details Standard DNA 1 (20 pM) 200 µL Standard DNA 2 (2 pM) 200 µL Standard DNA 3 (0.2 pM) 200 µL Standard DNA 4 (0.02 pM) 200 µL Standard DNA 5 (0.002 pM) 200 µL Standard DNA 6 (0.0002 pM) 200 µL 5x Primer Mix 2 mL (2 × 1 mL) 50x Dilution Buffer 1.7 mL Application Data Example 1.Comparison of PCR amplification efficiency of target genes with various GC contents Template DNAs of various GC contents bearing P5 and P7 adaptor sequences were analyzed. The GenNext™ NGS Library Quantification Kit achieved stable PCR efficiency with all targets, whereas other products showed poor specificity for targets with high GC contents as shown in the melting curve figure. Example 2.Comparison of PCR amplification efficiency of target genes with various fragment sizes Template DNAs of various fragment sizes (150–800 bp) bearing P5 and P7 adaptor sequences were analyzed. The GenNext™ NGS Library Quantification Kit achieved stable PCR efficiency with all targets, whereas other products showed poor efficiency with long targets. References Takagi M, Nishioka M, Kakihara H, Kitabayashi M, Inoue H, Kawakami B, Oka M, and Imanaka T., Appl. Environ. Microbiol., 63: 4504–10 (1997) Hashimoto H, Nishioka M, Fujiwara S, Takagi M, Imanaka T, Inoue T and Kai Y, J. Mol. Biol., 306: 469–77 (2001) Mizuguchi H, Nakatsuji M, Fujiwara S, Takagi M and Imanaka T, J Biochem., 126:762-8 (1999) Documents Download Manual Download SDS Request COA
