Amplify your cDNAs and your success with unsurpassed thermostability and sensitivity

 

Application

 

- First-strand synthesis of cDNA from RNA molecules (Reverse Transcription)
- RT-PCR
- Random priming reaction
- Library construction
- Probe labeling
- mRNA 5' end mapping by primer extension analysis
- Real time PCR

 

Experimental data

 

Figure 1. Comparison of amplification efficiency between AccuPower^® RocketScript™ Cycle RT PreMix and competitors’RTases

(a) Sensitivity test
Primer set: Human TFRC set
Lane M: 1 kb DNA Ladder
Lane1: 100 ng Human total RNA from HeLa cell
Lane2: 10 ng Human total RNA from HeLa cell
Lane3: 1 ng Human total RNA from HeLa cell
Lane4: 100 pg Human total RNA from HeLa cell
RT reaction condition is performed according to each manufacturer’s recommendations

(b) Full-Length cDNA synthesis test
RT reactions were performed according to each manufacturer’s recommendation. All cDNAs were amplified with AccuPower Hotstart PCR Premix (K-5050) from Bioneer
Note: Competitor products show inhibition at high input concentrations of total RNA
Lane 1: 1 μg Human total RNA from HeLa cell
Lane 2: 100 ng Human total RNA from HeLa cell
Lane 3: 10 ng Human total RNA from HeLa cell

 

 

Figure 2. Comparison of amplification quality between AccuPower Rocketscript™ Cycle RT PreMix(dT20) and other supplier's RT kit.
The reaction condition was performed according to each manufacturer's recommendation . All cDNAs were amplified with AccuPower 2X Greenstar qPCR Master mix (K-6251)from Bioneer.

 

 

Figure 3. Low copy species enrichment by cycle.
Comparing FTRT (Fixed Temperature Reverse Transcription) to 3 and 10 cycle(s) of CTRT reveal progressive improvement in detected cDNA yield as input copies decrease. FTRT: 30 min incubation at 42°C followed by 5 min deactivation at 95°C CTRT: Cycles of 15°C annealing 1min, 50°C cDNA synthesis 4 min, 60°C secondary structure melting and cDNA synthesis 20 sec.All cDNAs were amplified with AccuPower^® DualStar™ qPCR PreMix (K-6110) from Bioneer.

 

 

Figure 4. Comparison of amplification results between Bioneer RocketScriptTM Cycle RT PreMix and competitor RTases
All cDNAs were amplified with AccuPower Hotstart PCR Premix (K-5050) from Bioneer
Primer set: human Myc498 bp set
Lane M: 1 kb DNA Ladder
Lane 1: 10 ng Human total RNA from HeLa Cell
Lane 2: 1 ng Human total RNA from HeLa Cell
Lane 3: 100 pg Human total RNA from HeLa Cell
Lane 4: 10 pg Human total RNA from HeLa Cell

 

 

Figure 5. Highly reproducible Amplifications
Amplification of an 600bp target gene was detected using Human toal RNA(from 100ng to 10pg ) with AccuPower RocketScript™ cycle RT PreMix(dN12).
As shown in Fig5. Highly reproducible Amplifications were achieved within each Lot.
Lanes 1-5: 100 ng, 10 ng, 1 ng , 100pg and 10pg of total RNA from HeLa cells, respectively.

 

 

Specifications of Bioneer Enzymes