2X HoTaq™ One-Step Real-Time RT-PCR Kit For TaqMan®

Description:
RT-PCR is widely used for measuring gene expression in tissue samples or cell culture systems. Traditionally, it is performed in two separate reaction steps. First-strand cDNA is reverse-transcribed from total RNA or poly (A)+ RNA using a reverse transcriptase. Next, the cDNA is amplified by PCR using a DNA polymerase in another reaction.

The MCLAB's HoTaq Taqman probe One-step Real-time RT-PCR Kit offers a unique system for performing probe based realtime RT-PCR in a single step within a single tube with an optimized reaction condition, which utilizes our own proprietary engineered QuantumScript HD reverse transcriptase and hot-start Taq DNA Polymerase with high purity . No additional reagents or steps are required once the reaction is initialized. This novel kit enables to quantitatively detect specific RNA targets with high sensitivity, unparalleled convenience, and a wide dynamic range.

The technique reduces the risk of cross-contamination and minimizes the use of reagents. This method is particularly useful for applications in which the expression of a small number of genes that must be analyzed in many different total RNA samples, and robustly amplifing high-abundance transcripts from crude total RNA preparations.

Application:
- Gene Expression Analysis
- Genotyping
- Real-Time PCR

Primer and probe design: 
To achieve the best performance, appropriate software, such as ABI Primer Express^TM, should be used.

Recommended Reaction Conditions:
One cycle at 50°C for 10 to 30 minutes ^*;
One cycle at 95°C for 10 minutes;
Followed by 40 cycles of: 95°C for 15 seconds, 60°C for 1 minute ^#;
4°C hold (optional)
^* Reaction time could be adjusted according to RNA input.
^# Cycle number and annealing temperature are experiment dependent.

Recommended Storage Condition: -20°C

Notes:
To achieve accurate quantification, it is highly recommended to (1) avoid any RNase contamination; (2) design probe on sense strand; (3) use primer concentration from 100nM to 300nM; (4) shorten time between setting up reaction and loading plate onto PCR machine.

Advantages:
One-step RT-PCR products are faster.