THUNDERBIRD® Probe one-step qRT-PCR Kit, Trial (Only 1 Per Customer)
THUNDERBIRD® Probe one-step qRT-PCR Kit *Only 1 Per Customer*
A one-step real-time RT-PCR kit combining the highly efficient reverse transcriptase ReverTra Ace® with Tth DNA polymerase — for rapid, highly sensitive detection and quantification of small amounts of RNA.
Cold-chain shipping required. This product has ICE or DRY ICE shipping requirements — do not select Ground Shipping. The $1.00 listing applies to the sample only; please inquire. Only 1 Per Customer.
Tip: THUNDERBIRD is optimized for PCR amplicon size less than 500 bp and routine PCR purposes. Do not use it for PCR to amplify targets larger than 500 base pairs.
Product Overview
Ordering Information
Available configurations of this kit.
| Product Name | Cat. No. | Order |
|---|---|---|
| THUNDERBIRD® Probe one-step qRT-PCR Kit, Trial | TYB-QRZ-101S | Current page |
| THUNDERBIRD® Probe one-step qRT-PCR Kit, 1 kit | TYB-QRZ-101 | Order |
Description
THUNDERBIRD® Probe One-step qRT-PCR Kit is a one-step real-time reverse-transcription polymerase chain reaction (RT-PCR) kit that uses the highly efficient reverse transcriptase “ReverTra Ace®” and Tth DNA polymerase as a PCR enzyme. This product can be used mainly in TaqMan® probe assays. The one-step system is suitable for high-throughput analysis because of its simple reaction setup. In addition, this system can reduce the risk of cross-contamination. The combination of the two enzymes and optimized buffer system enable the effective detection and quantification of a small amount of RNA. This kit can also detect various kinds of RNA with different sequences because it is tolerant of target sequence diversity.
Features
Components
This kit includes the following components for 250 reactions, with 20 µL per reaction. All reagents should be stored at −20 °C.
| Solution | Volume |
|---|---|
| 2x Reaction Buff* | 2x 1.25 ml |
| DNA Polymerase | 125 µl |
| RT Enzyme Mix | 125 µl |
| 50x ROX Reference dye | 100 µl |
| RNase free water | 2x 1.25 ml |
Application Data
Example 1. The maximum sensitivities of various one-step qRT-PCR kits
A 4n dilution series of various viral RNAs was detected. The primers and TaqMan® probes were synthesized in accordance with previous reports. The graph indicates the minimum copy numbers that were detected by the kits. THUNDERBIRD® Probe One-step qRT-PCR Kit was the only kit that detected all viral RNAs tested at high sensitivity (≤30 copies).
Example 2. Comparison of sensitivity of detection of enterovirus RNA
The sensitivity and quantitativity of various kits were compared by detecting serially (4n) diluted enterovirus RNA. The primers and probe were synthesized in accordance with a previous report. Applied Biosystems® StepOnePlus™ was used in this experiment. THUNDERBIRD® Probe One-step qRT-PCR Kit was the only kit that detected less than 10 copies of RNA and showed wide-ranging quantitation. The results indicate that this kit is suitable for the highly sensitive detection of RNA viruses or mRNA expressed at a low level.
Example 3. Comparison of sensitivity of detection of enterovirus RNA
The expression levels of IL-1β, TNF-α and GAPDH mRNAs were analyzed using 10n times serially diluted total RNA (1 pg–100 ng) by triplex detection systems with TaqMan® probes labeled by different fluorescent dyes (Fig. 1). LightCycler® 96 (Roche Diagnostics) was used in this experiment. HeLa S3 cells were incubated for 20 h after being seeded in six-well plates at 4 × 105 cells/well and treated with or without 100 nM phorbol 12-myristate 13-acetate. Then, the expression levels of mRNA were analyzed using purified total RNA from treated cells. The elevations of IL-1β and TNF-α mRNAs were observed upon adding phorbol 12-myristate 13-acetate (Fig. 2). No significant differences of PCR efficiency and correlation coefficient were observed between the triplex and singleplex.
Example 4. SARS-CoV-2 Detection
Amplification using positive control RNA of the new coronavirus (SARS-CoV-2) showed amplification within 40 cycles when more than 50 copies of the positive control were used.
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.
