柏业贸易

特点及优势

灵活的条件
循环温度逆转录反应是通过反复进行2至3步以在高温（50至55℃）释放模板RNA二级结构并在低温（15至40℃）进行引物退火来进行的。可以比在42℃进行逆转录反应更高效率地合成CDNA。除循环温度逆转录反应外，还可以在22至55°C的单一温度下进行逆转录反应 .
稳定性
是一种包含稳定剂的干燥产品，由于提高了RTase的热稳定性，允许逆转录反应的温度高达55℃
反应时间可控
反应时间可以根据要扩增的基因的拷贝数和大小而变化。在高拷贝基因的情况下，可以通过10分钟的逆转录反应合成足够数量的cDNA
重现性
ISO 9001质量体系下的批量生产可最大程度地减少批次之间的差异，即使在重复实验中也可提供可重复的实验结果
便利性
添加了cDNA合成必不可少的所有成分（包括引物），只需添加RNA和D.W即可立即进行RT反应

应用

• 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

实验数据

cyclescript figure1

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

▶ Specifications

▶ Experimental data

cyclescript figure2

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)

cyclescript figure3

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

cyclescript figure4

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)

cyclescript figure5

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

cyclescript figure6

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.