Description

THUNDERBIRD® Probe qPCR Mix is a Taq DNA polymerase-based 2x Master Mix for highly efficient real-time PCR using TaqMan® probes. The master mix contains all required components, except for ROX reference dye, probe and primers. 50x ROX reference dye is individually supplied with this kit. The master mix facilitates reaction setup, and improves the reproducibility of experimental data.

This product is an improved version of Realtime PCR Master Mix (Code No. QPK-101) . In particular, reaction specificity and PCR efficiency is enhanced.

Features

• The master mix contains anti-Taq DNA polymerase antibodies for hot start technology. The specificity for the detection of low-copy targets is enhanced.
• The dispersion of PCR efficiency between targets is reduced by a new PCR enhancer (Patent pending) in the master mix.
• High specificity and robust enzyme enable the detection of a broad dynamic range.
• The reagent is applicable to most real-time cyclers (i.e. Block type and glass capillary type) including ABI, Bio-Rad, Roche, and TaKaRa. The 50x ROX reference dye in the kit can be applied to real-time cyclers that require a passive reference dye.

Specifications

• Hot Start
• Probe
• 2x Reaction mix
• 20µl total reaction volume
• Store at -20^oC
• 50x ROX reference dye

Components

• QPS-101T THUNDERBIRD® Probe qPCR Mix    1 ml × 1 50x ROX reference dye    50 µl × 1
•  
• QPS-101 THUNDERBIRD® Probe qPCR Mix    1.67 ml × 3 50x ROX reference dye    250 µl × 1
•  
• QPS-101X5 THUNDERBIRD® Probe qPCR Mix    (1.67 ml × 3) x 5 50x ROX reference dye    250 µl × 5

Compatible real-time instruments

Application data

Example 1. Comparison of the sensitivity and efficiency of the SYBR® Green I assay
β-actin mRNA was detected with serially diluted cDNA from HeLa cell total RNA. THUNDERBIRD® SYBR® qPCR Mix [Code No. QPS-201] showed greater sensitivity and efficiency than other kits (companies A and B).

Example 2. Comparison of the PCR efficiency
Norovirus G1 and human GAPDH genes were detected using serially diluted cDNA samples by SYBR® Green I and TaqMan real-time PCR. THUNDERBIRD® qPCR Master Mix showed greater efficiency than other reagents.

Detection of Norovirus G1 gene by SYBR® Green I assay. [Roche Diagnostics LightCycler 1.1]

Detection of GAPDH gene by TaqMan® assay. [Roche Diagnostics LightCycler 1.1]

Example 3. Verification of the measurement accuracy
Human GAPDH genes were detected using serially 20.5 fold diluted cDNA synthesised from HeLa cell total RNA by SYBR® Green I assay. THUNDERBIRD® qPCR Master Mix successfully detected the differences between dilutions.

Detection of human GAPDH gene by SYBR® GREEN I assay. [Applied Biosystems 7900HT]

Reference Cai, X., Warburton, C., Perez, O. F., Wang, Y., Ho, L., Finelli, C., ... & Meng, Z. (2024). Hippo-PKCζ-NFκB signaling axis: A druggable modulator of chondrocyte responses to mechanical stress . iScience , 27 (6), 109983. 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.

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