DESCRIPTION
THUNDERBIRD™ Probe and SYBR® qPCR Mix is a highly efficient 2x Master Mix for real-time PCR using TaqMan™ probes and SYBR® Green I. 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 experiments.
These products are improved versions of Realtime PCR Master Mix (Code No. QPK-101) and Realtime PCR Master Mix (Code No. QPK-201) . In particular, reaction specificity and PCR efficiency is enhanced.
This product is sold in U.S. under the license of US patent 7772383
from Chakrabarti Advanced Technology NewCo LLC.
FEATURES
- The specificity for the detection of low-copy targets is improved.
- The dispersion of PCR efficiency between targets is reduced by a new PCR enhancer*. (*Patent pending)
- High specificity and effective amplification 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). Because the 50x ROX reference dye is individually supplied with this kit, the kit can be applied to real-time cyclers that require a passive reference dye.
- The master mix contains anti-Taq DNA polymerase antibodies for hot start technology. The antibodies are easily inactivated in the first denaturation step, thereby activating the DNA polymerase.
STORAGE CONDITION
Store at -20ºC
COMPONENTS
| THUNDERBIRD™ Probe qPCR Mix | 1 mL × 1 |
| 50x ROX reference dye | 50 µL × 1 |
| THUNDERBIRD™ Probe qPCR Mix | 1.67 mL × 3 |
| 50x ROX reference dye | 250 µL × 1 |
| THUNDERBIRD™ SYBR® qPCR Mix | 1 mL × 1 |
| 50x ROX reference dye | 50 µL × 1 |
| THUNDERBIRD™ SYBR® qPCR Mix | 1.67 mL × 3 |
| 50x ROX reference dye | 250 µL × 1 |
COMPATIBLE REAL-TIME INSTRUMENTS
| Applied Biosystems |
ABI PRISM™ 7000 | Roche Diagnostics |
LightCycler™ 1.x / 2.0 |
|---|---|---|---|
| ABI PRISM™ 7700 | LightCycler™ Nano | ||
| Applied Biosystems™ 7300 | LightCycler™ 480 | ||
| Applied Biosystems™ 7500/7500FAST | Bio-Rad / MJ | MiniOpticon™ | |
| Applied Biosystems™ 7900HT | CFX96 Touch™ | ||
| Applied Biosystems™ StepOne™ | Agilent Technologies |
Mx3000P / Mx3005P / Mx4000 | |
| Applied Biosystems™ StepOnePlus™ | TaKaRa | Thermal Cycler Dice™ Real Time Systems |
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]