Description
The ExcelRT™ One-Step RT-qPCR kit (TaqMan, ROX) is designed for reverse transcription and quantitative real-time analysis of a specific target RNA by one-step reaction. The ExcelRT™ One-Step RT-qPCR kit (TaqMan, ROX), consisting of One-Step RT Enzyme Mix and 2X One-Step Master Mix, is a convenient kit designed for highly efficient cDNA synthesis and highly specific real-time PCR in a single tube. The One-Step RT Enzyme Mix contains a thermostable ExcelRT™ Reverse Transcriptase and a RNAok™ RNase inhibitor. Consequently, One-Step RT Enzyme Mix can reverse transcribe RNA to cDNA at a wide temperature range from 42 to 60°C and active against RNase A, RNase B and RNase C. By containing specialized hot-start Taq DNA polymerase, which greatly reduce primer-dimer formation and can be activated within 2 minutes, the 2X One-Step Master Mix features high specificity and is suitable for fast cycle program. This master mix includes ROX reference dye for normalization of each RT-qPCR assay.
Features
High yield
Reverse transcription at wide temperature range (42-60°C)
High specificity
Suitable for fast cycle program
With ROX reference dye
Storage
Aliquot to avoid multiple freeze-thaw cycles (stable within 30 freeze-thaw cycles)
Protect from light
-20°C for 12 months

Reverse transcription at wide template range
ExcelRT™ One-Step RT-qPCR Kit can quantitative analyze target RNA from a wide range of RNA template input. The amplification plot of one-step RT-qPCR with total RNA templates ranging from 1 pg to 1 ng in quantity, analyzed by using RQ2110 ExcelRT™ One-Step RT-qPCR Kit (TaqMan, ROX) for RT-qPCR amplification.

Reverse transcription at wide temperature range (42-60°C)
ExcelRT™ One-Step RT-qPCR Kit can quantitative analyze target RNA at a wide temperature range (42-60°C). The amplification plot of one-step RT-qPCR with reverse transcription at temperature range from 42 to 60°C, analyzed by using RQ2110 ExcelRT™ One-Step RT-qPCR Kit (A) or kit from brand N (B) for RT-qPCR amplification. The overlapped amplification curves display that ExcelRT™ One-Step RT-qPCR Kit preforms successfully cDNA synthesis at wide temperature range.
Contents
Component | Volume |
One-Step RT Enzyme Mix | 400 μl |
2X One-Step Master Mix | 2 x 1 ml |
Storage
Aliquot to avoid multiple freeze-thaw cycles (stable within 30 freeze-thaw cycles)
Protect from light
-20° for 12 months
Recommended primer design
Amplicon size: 80-150 bp
Tm value: around 60°C (calculated with Primer3 software)
Primer length: 17-25 mer
Sequence:
‒ 45-55% of GC content is recommended.
‒ Avoid regional high GC or AT content
‒ Avoid palindrome sequence
‒ Sequence with G or C at the 3' end is recommended.
Specificity of primers should be confirmed through a BLAST search.
Recommended probe design
Tm value: 6-10°C higher than primers
Probe length: 20-30 mer
Sequence:
‒ 35-65% of GC content is recommended.
‒ Avoid regional high GC or AT content
‒ Select the strand contains more C’s than G’s
‒ Avoid palindrome sequence
‒ Avoid a G at the 5' end to prevent quenching of the 5’ fluorophore.
Specificity of probe should be confirmed through a BLAST search.
Recommended reaction mixture set up for qPCR
Component | Volume | Final concentration |
Template RNA | Varied | 1 pg – 1 μg |
Forward primer (10 μM) | Varied | 125 – 900 nM |
Reverse primer (10 μM) | Varied | 125 – 900 nM |
TaqMan Probe (10 μM) | Varied | 100 – 200 nM |
One-Step RT Enzyme Mix | 2 μl | 1X |
2X One-Step Master Mix | 10 μl | 1X |
ddH2O | Up to 20 μl | - |
Total volume | 20 µl | - |
*Template amount varies depending on the copy number of target present in the template solution.
** The PCR primer and probe concentration for an optimal qPCR reaction may vary according to primers’ and probe’s properties.
Recommended qPCR Program
Standard program
Steps | Temp. | Time | Cycles |
Reverse transcription | 42°C - 60°C | 10 min | 1 |
Enzyme activation | 95°C | 3 min | 1 |
Denaturation | 95°C** | 15 sec | 40-50 |
Annealing/ Extension | 60°C | 1 min |
Fast program
Steps | Temp. | Time | Cycles |
Reverse transcription | 42°C - 60°C | 5 min | 1 |
Enzyme activation | 95°C | 20 sec | 1 |
Denaturation | 95°C** | 3 sec | 40-50 |
Annealing/ Extension | 60°C | 30 sec |
The effect of omega3 fatty acid supplementation on PPARγ and UCP2 expressions, resting energy expenditure, and appetite in athletes
Sara Moradi, Mohamadreza Alivand, Yaser KhajeBishak, Mohamad AsghariJafarabadi, Maedeh Alipour, Philip D. Chilibeck, and Beitullah Alipourcorresponding author, BMC Sports Sci Med Rehabil. 2021; 13: 48. Published online 2021 May 8. doi: 10.1186/s13102-021-00266-4
PMCID: PMC8106165
[RP1000] ExcelRT™ Reverse Transcriptase
High yield
Thermostable, up to 50°C, during first strand synthesis
High processivity, generating cDNA up to 8 kb
Reduced RNase H ribonuclease activity
[RP1400] ExcelRT™ Reverse Transcription Kit II
Contains all components for reverse transcription
High yield
Thermostable, up to 50°C
Reduced RNase H ribonuclease activity
Suitable for real-time PCR
Contains all components for reverse transcription
High yield
Thermostable, up to 50°C
Reduced RNase H ribonuclease activity
Suitable for real-time PCR
[TQ2110] ExcelTaq™ 2X Q-PCR Master Mix (TaqMan, ROX)
High sensitivity
High specificity
With ROX reference dye