Specifications

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment Size: Up to 10 kb

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
Lane 2: 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 Gentaur
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. Complex RNA amplification results of RocketScript Cycle RT PreMix
Each target gene was amplified after performing reverse transcription with AccuPower RocketScript Cycle RT PreMix.
Reverse transcription conditions: Conventional 1 hr incubation at 42°C, 50°C, or 60°C, deactivation at 95°C for 5 min
A: M-MLV Reverse Transcriptase
B: AccuPower RocketScript Cycle RT PreMix with Oligo (dT)20
Lane 1: 100 ng Human total RNA from HeLa cell
Lane 2: 10 ng Human total RNA from HeLa cell

Figure 3. Low copy species enrichment by cycle.
Comparing FTRT (Fixed Temperature Reverse Transcription) to 1 and 10 cycle(s) of CTRT reveal progressive improvement in detected cDNA yield as input copies decrease.

FTRT: 60 min incubation at 50°C followed by 5 min deactivation at 95°C
CTRT: Cycles of 37°C annealing 10 sec, 50°C cDNA synthesis 4 min, 55°C secondary structure melting and cDNA synthesis 
30 sec
Primer set: Human GAPDH
Human Total RNA from HeLa cells
qPCR with AccuPower GreenStar qPCR PreMix (K-6210)

Figure 4. Amplification comparison by cycle 

FTRT (Fixed Temperature Reverse Transcription) conditions:

CTRT (Cyclic Temperature Reverse Transcription) conditions:

Primer set: Human GAPDH and myc set
Lane M: 1 kb DNA Ladder
Lane1: 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. Comparison of amplification results between Gentaur RocketScript^TM Cycle RT PreMix and competitor RTases
All cDNAs were amplified with AccuPower Hotstart PCR Premix (K-5050) from Gentaur
Primer set: human Myc498 bp set
Lane M: 1 kb DNA Ladder
Lane1: 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

Specifications

5' to 3' exonuclease: No
3' to 5' exonuclease: No
3’ – A Overhang: No
Fragment Size: Up to 10 kb

Application

- First-strand synthesis of cDNA from RNA molecules (RT)
- RT-PCR
- Random priming reaction
- Library construction
- Probe labeling
- mRNA 5end Mapping by Primer Extension Analysis
- Real time PCR

Figure 1. Sensitivity comparison between AccuPower RocketScript RT PreMix and M-MLV RTase
Sensitivity results of AccuPower RocketScript RT PreMix using GAPDH compared with conventional Reverse transcriptases. 
Each 100 ng – 10 fg of total RNA used for RT and the same amount of RT products used for electrophoresis.
Lane 1: 10 fg Human total RNA from HeLa cell           Lane 2: 100 fg Human total RNA from HeLa cell
Lane 3: 1 pg Human total RNA from HeLa cell            Lane 4: 10 pg Human total RNA from HeLa cell
Lane 5: 100 pg Human total RNA from HeLa cell        Lane 6: 1 ng Human total RNA from HeLa cell
Lane 7: 10 ng Human total RNA from HeLa cell          Lane 8: 100 ng Human total RNA from HeLa cell

Figure 2. Comparison of amplification efficiency between AccuPower RocketScript RT PreMix and competitorsM-MLV RTase.
RocketScript is able to handle a wide range of sample concentrations and transcript lengths so your downstream applications are minimally effected by the reverse transcription step.
Lanes 1-3: 1,000 ng, 100 ng and 10 ng of total RNA from HeLa cells, respectively.

Figure 3. Sensitivity comparison between AccuPower RocketScript RT PreMix and competitor RTases using Real Time PCR
Reverse transcription conditions: conventional 1 hr incubation at 60°C, deactivation at 95°C for 5 min All cDNAs were amplified with AccuPower DualStar™ qPCR PreMix (K-6110) from Gentaur.

Concentration	RocketScript RT PreMix	Supplier Q	Supplier I
10,000	23.91	25.63	24.43
1,000	27.33	28.92	28.03
100	30.62	32.42	30.88
10	33.63	35.43	33.95
Efficiency	104%	103%	108%
Linearity	0.99999	0.9996	0.9995

Specifications

5' to 3' exonuclease activity: No
3' to 5' exonuclease activity: No
Terminal transferase activity: Yes
Fragment Size: Up to 10 kb

Application

Standard PCR

Figure 1. Processivity comparison with Lambda DNA template and different PCR reagents.

M: 100 bp Plus DNA Ladder (D-1035)
A: Accupower PCR PreMix
B: Taq mastermix from Company S
C: Taq mastermix from Company T
Lane M: 1 kb DNA Ladder (D-1040)
Lane 1: 1 kb PCR product
Lane 2: 2 kb PCR product
Lane 3: 3 kb PCR product
Lane 4: 4 kb PCR product

Figure 2. Comparison of sensitivity test for AccuPower PreMix PCR master mix and other companies' products using serial diluted human genomic DNA.

Reaction condition: 95°C for 5 minutes, followed by 35 cycles of 20 seconds at 95°C, 20 seconds at 60°C, 30 seconds at 72°C.

A: human Insulin receptor gene
B: DNA cross-link repair 1A gene
I: AccuPower PCR PreMix
II: Taq DNA Polymerase from Company A
III: Taq DNA Polymerase from Company Q
IV: PCR PreMix from Company I
M: 100 bp DNA ladder (D-1030)
Lane 1: 10 ng
Lane 2: 1 ng
Lane 3: 100 ng
Lane 4: 10 ng