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 QUANTIFICATION
    KOD SYBR® qPCR Mix can efficiently amplify GC- and AT-rich fragments of different lengths without bias.
  • BROAD DYNAMIC RANGE
    The kit has a broad dynamic range from 20 pM (Standard DNA 1) to 0.0002 pM (Standard DNA 6).
  • CONVENIENt
    The 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.

APPLICATIONS

  • NGS Library quantification for illumina’s insturment (ex. MiniSeq, MiSeq, HiSeq, NextSeq)

STORAGE CONDITION

Store at -20℃

COMPONENTS

< KOD SYBR® qPCR mix* >
KOD SYBR® qPCR mix 1.67 ml x 3
50x ROX reference dye 250 µl
< Standard & Primer Set >
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)