Robustart Tth

Robustart Tth is a hot-start Tth enzyme developed by our company. It not only has 5 '-3' DNA template-dependent polymerase activity but can also be widely applied in various PCR reactions. It also has higher high-temperature reverse transcription activity in the presence of Mn²+, which can effectively eliminate the adverse effects of RNA advanced structure and non-specific annealing of primers at low temperatures on cDNA synthesis, thereby increasing the specificity of reverse transcription, improving the efficiency and yield of amplification, and can be used for single-enzyme one-step RT-PCR reactions. Robustart Tth has its activity blocked at room temperature and better specificity, which can significantly enhance the amplification effect of low-concentration templates.

1．5 U/µL Robustart Tth 

2．10×Tth RT Buffer Basic (Mn²+ free)（选配） 

3.25mM Mn(OAc)₂ (optional)

*10×Tth RT Buffer Basic (Mn²+ free) does not contain DNNTP and Mn²+. Please add dNTPs and Mn(OAc)₂ when preparing the reaction system.

Antibody modification, above 90℃, hot start for 30 seconds to 1 minute.

2. It has dual functional activities of polymerase and reverse transcriptase.

3. In the presence of Mn²+, reverse transcription can be carried out at 60℃-70℃, and the reverse transcription efficiency of complex RNA templates is higher, reducing non-specificity.

4. Good tolerance to inhibitors.

1.Quantitative PCR.

2.Multiplex PCR, genotyping detectionHigh-sensitivity virus detection.

1. Freeze-thaw stability verification

Experimental conditions

Experimental results

Test method: Freeze and thaw a single enzyme for 5, 10, 15, and 20 times. Amplification tests were conducted in the MD2011 system with 0 freeze-thaw cycles as the control.

Summary: In the human genome, the amplification performance of the E17 single enzyme was consistent with that of the unthawed reagent after 5, 10, 15, and 20 freeze-thaw cycles, and it could remain stable.

2. Accelerate stability verification

Experimental conditions

Experimental results

Test method: Accelerate the single enzyme for 7 and 14 days. Amplification tests were conducted in the MD2011 system with 0-day acceleration as the control.

Summary: In human genome detection, after E17 single enzyme was placed at 37℃ and accelerated for 7 days and 14 days, its amplification performance was consistent with that of the unaccelerated reagent and could remain stable.

3. Verification of transportation stability

Experimental conditions

Experimental results

Test method: After transporting the single enzyme by SF Express Air for 84 hours, the reagent stored at -20℃±5℃ was used as the control. Verification is carried out in the MD2031 system

Summary: After 84 hours of transportation, E17 samples of the same batch were stored at -20°C±5° C. Under the human genome template diluted with human whole blood, the differences in ΔCT values at each channel were all less than 0.5. Rn% is less than 10%, and the linear fitting degree is good.

4. Anti-inhibition verification (Oral swab direct expansion test)

Experimental conditions

Experimental results

Test method: Scrape the left and right sides of the inner wall of the mouth 10 times with a cotton swab, then elute in 1 mL H2O, centrifuge at 2500 r/min for 10 minutes, remove the supernatant, and leave about 100 μL. Mix well and add the sample directly.

Summary: E17 can directly amplify oral swabs, with normal Ct values and a good amplification curve.