AccuPower® RocketScript™ RT Master Mix, RNase H Minus (2X, 1ml)

AccuPower^® RocketScript™ RT Master Mix, RNase H Minus is the Master Mix type of AccuPower^® RocketScript™ RT Premix, RNase H Minus products.

※ Master Mix product is shipped in dry ice

$239.00

Cat. No.

K-2249

Product Overview

Features and Benefits

Rapid Synthesis Short-Time Synthesis : 9 kb synthesis possible with 10 minute RT reaction
Time-efficient and economical synthesis of cDNA from secondary structure RNA with RocketScript™ RNase having excellent thermal stability capable of being active at even 70 ℃
No RNase H activity
Useful for cDNA synthesis up to 12.5 kb from long target RNA using enzymes developed by proprietary genetic engineering technology
Sensitivity
High sensitivity and effective synthesis of cDNA even at scarce amount of 1 pg of human total RNA.
Convienance
Synthesis of cDNA samples difficult to be done using traditional methods, such as those made with long size RNA (9 kb), possible only within 10 minutes
Reproducibility
Minimized deviation between lots, then supplied as uniform quality product to provide reproducible experimental results by mass production under ISO 9001 quality system
Application

• First-strand synthesis of cDNA from RNA molecules (RT)

• Random priming reaction

• cDNA library construction

• Probe labeling

• mRNA 5‘-end mapping by primer extension analysis

• PCR

• Real-time qPCR

Experimental data

Figure. Performance of the AccuPower^® RocketScript ™ RT PreMix, RNase H Minus in two-step RT-PCR.
Target mRNAs ranged from 500 bp to 11 kb were reverse transcribed by using AccuPower^® RocketScript ™ RT PreMix, RNase H Minus. With each cDNA, PCR was performed by using AccuPower^® ProFi Taq PCR PreMix (Cat. No. K-2631, Bioneer). Template sizes are indicated above the gel image.

M1; 1 kb DNA Ladder (Cat. No. D-1040, Bioneer)
M2; Lambda DNA/Hind III Markers (Cat. No. D-1050, Bioneer)

Technical/Specs

▶ Specifications

DNase activity	No
RNase activity	No
RNase H activity	No
Fragment size	Up to 12.5 kb

▶ Experimental data

Figure 1. Comparison of cDNA synthesis efficiency of AccuPower^® RocketScript ™ RT PreMix, RNase H Minus and other suppliers’ products.
Synthesis of cDNA was conducted at 50℃ or 42℃ for 10, 20, 30 and 60 min with 1 µg of 3.4 kb to 9 kb RNA transcript (total RNA from HeLa cells) as a template by using AccuPower^® RocketScript ™ RT PreMix, RNase H Minus and other kits from competitors. With each cDNA, PCR was performed by using AccuPower^® ProFi Taq PCR PreMix
(Cat. No. K-2631, Bioneer). PCR products were analyzed on a 1% alkaline agarose gel.

Lane 1; RT 10 min Lane 2: RT 20 min
Lane 3; RT 30 min Lane 4: RT 60 min
M; 1 kb DNA Ladder (Cat. No. D-1040, Bioneer)

Figure 2. Thermal stability of AccuPower^® RocketScript ™ RT PreMix, RNase H Minus.
Reverse transcription using AccuPower^® RocketScript ™ RT PreMix, RNase H Minus was performed at 50℃ to 70℃ for 30 min with 100 ng, 10 ng, 1 ng and 100 pg of 600 bp RNA transcript as a template. Even at 70℃, AccuPower^® RocketScript ™ RT PreMix, RNase H Minus successfully completed cDNA synthesis. With each cDNA, PCR was performed by using AccuPower^® PyroHotStart Taq PCR PreMix (Cat. No K-2611, Bioneer).

Lane 1; 100 ng of total RNA from HeLa cells
Lane 2; 10 ng of total RNA from HeLa cells
Lane 3; 1 ng of total RNA from HeLa cells
Lane 4; 100 pg of total RNA from HeLa cells
M; 100 bp DNA Ladder (Cat. No. D-1030, Bioneer)

Figure 3. Sensitivity comparison of RTase products with various targets.
Various target mRNAs were reverse transcribed at 50℃ for 30 min by using AccuPower^® RocketScript™ RT PreMix, RNase H Minus and competitor’s product, respectively. With each cDNA, PCR was performed by using AccuPower^® PyroHotStart Taq PCR PreMix (Cat. No K-2611, Bioneer).

Lane 1; 10 ng of total RNA from HeLa cells
Lane 2; 1 ng of total RNA from HeLa cells
Lane 3; 100 pg of total RNA from HeLa cells
Lane 4; 10 pg of total RNA from HeLa cells
M; 100 bp DNA Ladder (Cat. No. D-1030, Bioneer)

Figure 4. Broad dynamic range of AccuPower^® RocketScript ™ RT PreMix, RNase H Minus.
First-strand cDNA was synthesized using the AccuPower^® RocketScript ™ RT PreMix, RNase H Minus. cDNA was amplified using the AccuPower^® Plus DualStar™ qPCR PreMix (Cat. No. K-6600, Bioneer) on the Exicycler™ 96 Real-Time Quantitative Thermal Block from Bioneer. The standard curve illustrates high linearity (R2 = 0.999) across a broad range of input RNA, suggesting that the relative representation of specific RNA transcripts is preserved in the cDNA pool regardless of the abundance of total RNA. Amplification was performed on 10-fold serial dilutions of HeLa total RNA (1 μg to 1 pg) and mouse total RNA (10 μg to 1 pg).

Literature/Support

◈ Quality Assurance

Bioneer is the holder of Quality Management System Certificates for the following standards.

• ISO 9001 - certificate

Ordering Info

Cat. No.	Product Description	Option				Price
K-2221	AccuPower^® RocketScript™ RT PreMix & Master Mix, RNase H Minus	PreMix type	0.2 ml thin-wall 8-tube strips with attached cap / 96 tubes		20 µl/rxn	$K-2221
K-2223					50 µl/rxn	$K-2223
K-2245				dN6	20 µl/rxn	$K-2245
K-2246				dN6	50 µl/rxn	$K-2246
K-2247				dN12	20 µl/rxn	$K-2247
K-2248				dN12	50 µl/rxn	$K-2248
K-2241				dT20	20 µl/rxn	$K-2241
K-2243				dT20	50 µl/rxn	$K-2243
K-2222			0.2 ml thin-wall 8-tube strips with attached cap / 480 tubes		20 µl/rxn	$K-2222
K-2224					50 µl/rxn	$K-2224
K-2242				dT20	20 µl/rxn	$K-2242
K-2244				dT20	50 µl/rxn	$K-2244
K-2249*		MasterMix type	1 ml of 2X Master mix solution			$K-2249

*For MasterMix products, Primers(dT20&dN6) are provided in a separate tube.

AccuPower® RocketScript™ RT Master Mix, RNase H Minus (2X, 1ml)

Features and Benefits

Experimental data

▶ Specifications

▶ Experimental data

◈ Manual

◈ MSDS

◈ Brochure

◈ Quality Assurance