GenNext NGS Library Quantification Kit, 1 kit
GenNext NGS Library Quantification Kit, 1 kit
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 adapt…
| Product Name | Cat. No. | Size | Storage | Order |
|---|---|---|---|---|
| GenNext NGS Library Quantification Kit, Trial | TYB-NLQ-101S | — | -20°C | Order |
| GenNext NGS Library Quantification Kit, 1 kit | TYB-NLQ-101 | — | -20°C | Current page |
GenNext NGS Library Quantification Kit
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.
Application
NGS Library quantification for illumina’s insturment (ex. MiniSeq, MiSeq, HiSeq, NextSeq)
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
Applications 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)
REAL-TIME PCR KITS
DiagnoCine offers TOYOBO® Taq DNA Polymerases in the form of Master Mix (Probe), SYBR Green Master Mix (Plus), Thunderbird qPCR (Probe), and Thunderbird SYBR qPCR formats. All Taq DNA Polymerases will provide the highest specificity by Hot Start technology, excellent Passive reference, and fits well for glass capillaries for many purposes.
• Real-time PCR Master Mix is a probe assay type with the highest hot start specificity, fast routine PCR experiments, and works well for GC-rich regions. This is for routine short-length PCR targets and not suitable for the long target (>10kb length; KOD is best for long-range size PCR) PCR. It can use for crude samples but KOD is best for crude sample amplifications. This product's strongest feature is the high specificity and Probe master mix for routine end-point PCR experiments.
• SYBR Green Realtime PCR Master Mix (plus for a hot start) is not as specific as the other Taq polymerases (i.e. Probe type or Thunderbird SYBR are more specific) master mix but this brand provides an excellent passive reference and fits for glass capillary use. It is designed to work well for routine short-range PCR and light GC-rich regions and fairly clean crude samples.
• Thunderbird Probe qPCR Mix is a probe assay type with hot start specificity, good passive reference, and glass capillary type PCR experiments. The strength is the highest specificity with low noise bands for difficult PCR and routine experiments such as genotyping and colony screening. Promised for best specificity and fast PCR speed but not made for a long-range PCR (>10kb length; KOD is best for long amplicons more than 10kb). It can unwind the GC-rich region and crude samples can be used (KOD is the best for crude sample use). This type is best for all routine laboratory PCR with short-mid range amplicons, looking for fast PCR time but with the strongest specificity and not dealing with PCR noise bands.
• Thunderbird SYBR qPCR Mix is an SYBR assay type with a strong specificity but slightly less specific than Probe qPCR. It has passive reference and glass capillary compatibility. The product is designed for a routine PCR with short to mid-range amplicons for a short and fast PCR time. The best performance is expected with partially clean crude samples and a fair level of GC-rich complexity.
