AccuPower® CycleScript RT Master Mix (2X, 1ml)

AccuPower^® CycleScript™ RT MasterMix is the Master Mix type of AccuPower^® CycleScript™ RT Premix products.

※ Master Mix product is shipped in dry ice

$149.00

Cat. No.

K-2051

Product Overview

Features and Benefits

Flexible conditions
CRCT (Cyclic Temperature Reverse Transcription) performing a primer annealing stage at a range of low temperature from 15 to 40℃) and the release of secondary structure of template DNA by repeating the step 2 or 3 times at a range of high temperature from 50 to 55°C, allowing more efficient synthesis of cDNA with higher temperature than the traditional methods undergoing the process at 42°C .
Stability
Strong thermal stability of RTase with the use of dried products containing a stabilizer allowing to react at a higher temperature up to 55℃
Controllable reaction time
Varied reaction time depending on the number and size of copies of the gene to be amplified, with synthesis high-copy gene cDNA within 10 minutes through reverse transcription reaction.
Reproducibility
Minimized deviation between lots and reproducible results even for replicates tested repeatedly under same conditions and variables by mass production under ISO 9001 quality system
Easy of use
Simplified procedures with all components essential for cDNA synthesis, including the primers, added in the kit to perform the RT reaction immediately just by adding the RNA to be amplified and D.W.

Application

• Sequencing single and double-strand DNA or RNA

• Random priming reaction

• cDNA library construction

• Probe labeling

• mRNA 5'-end mapping by primer extension analysis

• PCR

• Real-time PCR

Experimental data

Figure 1. Reaction conditions of Cyclic RT compared with that of conventional RT.

The conventional fixed temperature RT reactions at 42°C and 55°C and cyclic RT reactions at 37°C and 50°C using HeLa cell total RNA (800 ng each) were performed 800 bp, 1.5 kb, 2.0 kb, and 2.6 kb fragments of human transferrin receptor gene were amplified with each primers. AccuPower^® CycleScript RT Premix series (dT20 & dN12) preferably shows good results at cyclic RT reaction condition.

Technical/Specs

▶ Specifications

DNase activity	No
RNase activity	No
Fragment size	Up to 9 kb

▶ Experimental data

Figure 1. Amplification Results of AccuPower^® CycleScript RT Premix series (dT20 & dN12) using Coxsaekie virus A9 (AJ295200) 5'-UTR region compared with other companies.
RT reactions were performed according to manufacturer's recommendation with Coxaekie virus RNA. PCR reactions were performed AccuPower^®PCR PreMix (K-2012) from Bioneer.

1; AccuPower RT Premix RT reacted with dT primer
2; AccuPower RT Premix RT reacted with random primer
3,4; AccuPower CycleScript RT PreMix (dT20)
5,6; AccuPower CycleScript RT PreMix (dN12)
7,8; AccuPower CycleScript RT PreMix (dT20)
9,10; AccuPower CycleScript RT PreMix (dN12)
11,12; AccuPower CycleScript RT PreMix (dT20)
13,14; AccuPower CycleScript RT PreMix (dN12)
15,16; C company's RT product with dT primer RT
17,18; C company's RT product with random primer RT
19,20; I company's RT product with dT primer RT
21,22; I company's RT product with random primer RT
M; 100 bp ladder (Cat. No.: D-1030)

Figure 2. Synthesis test of long transcript compared with other companies.
Bioneer's AccuPower^® CycleScript RT PreMix 1 and 2 were incubated for 1 hr, at 55°C, C company's RT Product 3 and 4 were for 1 hr, at 42°C, I company's RT product 5 and 6 were for 1 hr, at 45°C. And then 95°C 5 min with HeLa cell total RNA. RT reaction condition is performed according to each manufacturer's recommendation.

M; 100 bp ladder (Cat. No.: D-1030) 1, 3, 5; RT product included dT Primer
2, 4, 6; RT product included random primer

Figure 3. Gene amplification test of transferrin receptor compared with other companies.
The reaction condition was performed according to each manufacturer's recommendation. All cDNAs were reacted with AccuPower^® PCR PreMix (K-2012) from Bioneer

1~ 5; AccuPower^® CycleScript RT PreMix (dT20) incubated at 55°C for 1 hr
6~10; AccuPower^® CycleScript RT PreMix (dN12) incubated at 55°C for 1 hr
11~15; C company's RT product including dT primer incubated at 42°C for 1 hr
16~20; C company's RT product including random primer incubated at 42°C for 1 hr
21~25; I company's RT product including dT primer incubated at 45°C for 1 hr
26~30; I company's RT product including random primer incubated at 45°C for 1 hr
M; 1 kb ladder (Cat. No.: D-1040)

Figure 4. RT reaction at various temperature and short reaction time
Reverse transcription condition: conventional 42°C 1 hr, 55°C 1 hr & Cyclic reaction 1: (37°C 2 min/50°C 3 min) X 12, Cyclic reaction 2 : (37°C 1 min/ 47°C 3 min/55°C1 min) X 12.
This product shows thermal stability.

Primer set : human GAPDH primer set, human β-actin primer set M: 100 bp ladder (D-1030)
1,5,9,13,17,21,25,29; HeLa cell total RNA 100 ng 2,6,10,14,18,22,26,30; HeLa cell total RNA 10 ng
3,7,11,15,19,23,27,31; HeLa cell total RNA 1 ng 4,8,12,16,20,24,28,32; HeLa cell total RNA 100 pg

Figure 5. AccuPower^® CycleScript RT Premix (dT20) was performed cyclic reverse transcription according to (37°C 2 min/50°C 3 min) ; X12, X4, X2 cycles. 10min reaction is enough. HeLa cell total RNA templates are serial dilution such as 100 ng, 10 ng, 1 ng, 100 pg.

Literature/Support

◈ Quality Assurance

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

• ISO 9001 - certificate

• ISO 13485 - certificate

Ordering Info

Cat. No.	Product Description	Option				Price
K-2046	AccuPower^® CycleScript™ RT PreMix & Master Mix	PreMix type	0.2 ml thin-wall 8-strip tubes with attached cap / 96 tubes	20 µl reaction	dN6	$K-2046
K-2045					dN12	$K-2045
K-2044					dT20	$K-2044
K-2049				50 µl reaction	dN6	$K-2049
K-2048					dN12	$K-2048
K-2047					dT20	$K-2047
K-2046-B			0.2 ml thin-wall 8-strip tubes with attached cap / 480 tubes	20 µl reaction	dN6	$K-2046-B
K-2045-B					dN12	$K-2045-B
K-2044-B					dT20	$K-2044-B
K-2049-B				50 µl reaction	dN6	$K-2049-B
K-2048-B					dN12	$K-2048-B
K-2047-B					dT20	$K-2047-B
K-2050-2			0.5 ml thin-wall tubes with attached cap / 100 tubes	50 µl reaction	dN6	$K-2050-2
K-2050-1					dN12	$K-2050-1
K-2050					dT20	$K-2050
K-2051		MasterMix type	1 ml of 2X Master mix solution			$K-2051

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