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商品详细Qiagen/QIAGEN OneStep RT-PCR Kit/For 25 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 50 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 250 µl), dNTP Mix (1 x 50 µl, 10 mM each), 5x Q-Solution (1 x 400 µl), RNase-Free Water (1 x 1.9 ml)/210210

Qiagen/QIAGEN OneStep RT-PCR Kit/For 25 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 50 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 250 µl), dNTP Mix (1 x 50 µl, 10 mM each), 5x Q-Solution (1 x 400 µl), RNase-Free Water (1 x 1.9 ml)/210210

商品编号： 210210

品牌： Qiagen

市场价： ¥3420.00

美元价： 2052.00

产地：美国(厂家直采)

公司：

产品分类：多肽合成

公司分类： peptide

联系Q Q： 3392242852

电话号码： 4000-520-616

电子邮箱： info@ebiomall.com

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商品介绍

PCR Enzymes & Kits
End-Point PCR17
HiFidelity, Long-Range & Other PCR6
One-Step RT-PCR2
Reverse Transcription & cDNA Synthesis2

QIAGEN OneStep RT-PCR Kit

For highly sensitive andspecific one-step RT-PCR

Fast and easy one-tube setup
Efficient one-step RT-PCR of any RNA template without optimization
Unique enzyme mix for high specificity and sensitivity
Balanced mixture of enzymes with optimized reverse-transcription buffer

The QIAGEN One-Step RT-PCR Kit provides a blend of Sensiscript and Omniscript Reverse Transcriptases, HotStarTaq DNA Polymerase, QIAGEN OneStep RT-PCR Buffer, a dNTP mix, and Q-Solution, a novel additive that enables efficient amplification of "difficult" (e.g., GC-rich) templates. The easy one-tube setup and optimized components result in highly sensitive and successful results.

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Product Details
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Product Resources

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Cat No./ID:210210

QIAGEN OneStep RT-PCR Kit (25)

$171.00

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For 25 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 50 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 250 µl), dNTP Mix (1 x 50 µl, 10 mM each), 5x Q-Solution (1 x 400 µl), RNase-Free Water (1 x 1.9 ml)

Cat No./ID:210212

QIAGEN OneStep RT-PCR Kit (100)

$615.00

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For 100 x 50 µl reactions: QIAGEN OneStep RT-PCR Enzyme Mix (1 x 200 µl), 5x QIAGEN OneStep RT-PCR Buffer (1 x 1 ml), dNTP Mix (1 x 200 µl, 10 mM each), 5x Q-Solution (1 x 2 ml), RNase-Free Water (2 x 1.9 ml)

Cat No./ID:210215

QIAGEN OneStep RT-PCR Kit (1000)

$5,355.00

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For 1000 x 50 µl reactions (available in a single tube): QIAGEN OneStep RT-PCR Enzyme Mix (2 x 1 ml), 5x QIAGEN OneStep RT-PCR Buffer (1 x 10 ml), dNTP Mix (2 x 1 ml, 10 mM each), 5x Q-Solution (1 x 10 ml), RNase-Free Water (1 x 40 ml)

The QIAGEN OneStep RT-PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

Increased specific primer annealing.

Ammonium and potassium cations in QIAGEN PCR Buffers increase specificity of primer annealing. K⁺binds to the phosphate groups (P^–) on the DNA backbone, stabilizing the annealing of the primers to the template. NH₄⁺, which exists both as the ammonium ion and as ammonia under thermal-cycling conditions, can interact with the hydrogen bonds between the bases (B), destabilizing the weak hydrogen bonds at mismatched bases. The combined effect of the two cations maintains a high ratio of specific-to-nonspecific primer-template binding over a wide temperature range.

Efficient detection of viral RNA.

A 336 bp fragment of F-gene mRNA was reverse-transcribed and amplified from Sendai virus RNA isolated from persistently infected Vero cells. Reactions were prepared using the QIAGEN OneStep RT-PCR Kit and the indicated number of viral genome copies. M: markers. (Data kindly provided by H. Rausch, Max Planck Institute for Biochemistry, Martinsried, Germany as part of the project "Experimental control of virological work at safety levels 2 and 3 in Bavaria," supported by the Bavarian Ministry of the Environment.)

RT-PCR of GC-rich template.

A 439 bp fragment of transcription factor TAFII100 mRNA (GC content: 75%) was reverse-transcribed and amplified from HeLa-cell total RNA. Reactions were prepared using the QIAGEN OneStep RT-PCR Kit with (+) or without (–) Q-Solution. Q-Solution ensures specific amplification of GC-rich templates. M: markers.

Highly specific RT-PCR using low amounts of template.

One-step RT-PCR was carried out using the indicated amounts of total RNA from HeLa cells and primers specific for α-catenin, amplifying a 690 bp product. All reactions were carried out following suppliers" instructions. M: 100 bp ladder.

Influence of annealing temperature on specificity.

One-step RT-PCR was performed using kits from the indicated suppliers over a range of annealing temperatures. A 1289 bp fragment from the human RCC1 gene was reverse transcribed and amplified from Hela RNA (arrow). M: markers. High levels of specific amplification without optimization were observed only with the QIAGEN OneStep RT-PCR Kit.

OneStep RT-PCR procedure.

The QIAGEN OneStep RT-PCR Kit allows fast and easy RT-PCR setup. Whatever the application — virus detection, molecular diagnostics research, or gene expression analysis— just mix all components together in one tube and start the thermal-cycler program. The reaction mixture contains all of the reagents required for both reverse transcription and PCR; nothing needs to be added once the reaction has been started.

Performance

The QIAGEN OneStep RT-PCR Kit provides a convenient format for highly sensitive and specific RT-PCR using any RNA template. The kit includes optimized components that allow both reverse transcription and PCR amplification to take place in the same reaction mix in a "one-step" reaction. A unique enzyme combination and specially developed reaction buffer ensure efficient, highly specific reverse transcription and PCR in one tube, without the need for optimization (see figures "Highly specific RT-PCR using low amounts of template" and "Efficient detection of viral RNA"). The innovative, dual-cation PCR buffer provided with the kit ensures high yields of specific PCR product over a wide range of annealing temperatures (see figure "Influence of annealing temperature on specificity"). Suboptimal RT-PCR is improved using Q-Solution, a unique additive that facilitates reverse transcription and amplification of templates with a high GC content or a high degree of secondary structure (see figure "RT-PCR of GC-rich template").

Principle

The QIAGEN OneStep RT-PCR Kit is designed for easy and sensitive one-step RT-PCR using any RNA template.

The QIAGEN OneStep RT-PCR Enzyme Mix contains a specially formulated enzyme blend for both reverse transcription and PCR. The unique combination of Omniscript and Sensiscript Reverse Transcriptases, with their high affinity for RNA templates, ensures highly efficient and sensitive transcription of RNA amounts from as little as 1 pg up to 2 µg. After reverse transcription, reactions are heated to 95°C for 15 minutes to activate HotStarTaq DNA Polymerase and simultaneously inactivate the reverse transcriptases. This hot-start step eliminates nonspecific amplification products such as primer dimers and reduces background smear, ensuring highly sensitive and reproducible RT-PCR (see figures "Highly specific RT-PCR using low amounts of template" and "Efficient detection of viral RNA").

Wide range of annealing temperatures

The optimal primer annealing temperature is dependent on the base composition (i.e., the proportion of A, T, G, and C nucleotides), primer concentration, and ionic reaction environment. QIAGEN PCR Buffers contain both K⁺ and NH₄⁺ and deliver high yields of specific PCR product over a wide range of annealing temperatures (see figure "Increased specific primer annealing"). This specificity is achieved by destabilizing non-specifically bound primers, providing a more robust reaction environment and eliminating the need for tedious annealing temperature optimization. In contrast, the range of optimal PCR annealing temperatures is smaller and less predictable using a PCR or one-step RT-PCR buffer that only contains K⁺, as illustrated in figure "Influence of annealing temperature on specificity".

QIAGEN OneStep RT-PCR Buffer has been specially developed to allow both efficient reverse transcription and PCR amplification. The buffer contains novel additives that prevent inhibition of PCR amplification by reverse transcriptases, a problem often encountered in one-step RT-PCR. The buffer ensures specific primer annealing over a wide range of temperatures and Mg²⁺ concentrations; providing robust and highly efficient RT-PCR from any RNA template. The QIAGEN OneStep RT-PCR Kit includes Q-Solution, an innovative additive that modifies the melting behavior of nucleic acids. Q-Solution facilitates reverse transcription and amplification of templates with a high GC content or a high degree of secondary structure (see figure "RT-PCR of GC-rich template"). Using Q-Solution simplifies optimization of RT-PCR for difficult templates. The QIAGEN OneStep RT-PCR Kit includes everything required for faster and easier RT-PCR for even the most sensitive applications (see table).

Reliable one-step RT-PCR results
QIAGEN OneStep RT-PCR Kit contents	Features
HotStarTaq DNA Polymerase	Highly specific products
Sensiscript and Omniscript Reverse Transcriptases	Wide range of RNA amounts (1 pg – 2 µg) High sensitivity
OneStep RT-PCR Buffer	Minimal optimization needed No inhibition of PCR by reverse transcriptasesInhibition of RNases
Q-Solution	Facilitates amplification of GC-rich templates

Procedure

The QIAGEN OneStep RT-PCR Kit allows fast and easy RT-PCR setup. Whatever the application — virus detection, molecular diagnostics research, or gene expression analysis — just mix all components together in one tube and start the thermal-cycler program (see flowchart "OneStep RT-PCR procedure"). The reaction mixture contains all of the reagents required for both reverse transcription and PCR; nothing needs to be added once the reaction has been started (see table).

Applications

The QIAGEN OneStep RT-PCR Kit is suitable for RT-PCR applications such as:

Virus detection
Single-cell RT-PCR
Gene-expression analysis

Specifications

Features	Specifications
Applications	Gene-expression analysis, virus detection
Enzyme activity	Reverse transcription, 5" -> 3" exonuclease activity
Mastermix	No
Reaction type	One-step RT-PCR
Real-time or endpoint	Endpoint
Sample/target type	RNA template
Single or multiplex	Single
With/without hotstart	With hotstart

Product Resources

Brochures & Guides (3)

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	Analyzing Genetic Differences - (EN) Second edition — innovative tools	Show details
	(EN) - Maximizing PCR and RT-PCR success — Third Edition Addressing critical factors and new solutions	Show details
	iPP QIAGEN OneStep Ahead RT-PCR Kit	Show details

(1)

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	Product Profile - OneStep RT-PCR Kit	Show details

FAQs (7)

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	Is mRNA isolation necessary for sensitive RT-PCR? FAQ ID -111	View
	Do I need to use an RNase inhibitor in my RT reaction? FAQ ID -119	View
	Does QIAGEN sell Q-Solution separately? FAQ ID -204	View
	Does the 5x OneStep RT-PCR Buffer contain BSA? FAQ ID -326	View
	Can I shorten the activation time for the HotStarTaq DNA Polymerase? FAQ ID -565	View
	Are there specific recommendations for performing RT-PCR on RNA isolated from paraffin-embedded samples? FAQ ID -828	View
	Do you have a protocol for polyacrylamide gel analysis of oligonucleotides? FAQ ID -961	View

Kit Handbooks (1)

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	(EN) - QIAGEN OneStep RT-PCR Kit Handbook For fast and efficient one-step RT-PCR	Show details

Technical Information (1)

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	(EN) - Maximizing end-point PCR success with QIAGEN"s automatable PCR solutions	Show details

Supplementary Protocols (4)

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	Using the QIAGEN® OneStep RT-PCR Kit with a 25 µl reaction volume - (EN)	Show details
	Using the QIAGEN® OneStep RT-PCR Kit and Q-Solution™ with a 25 µl reaction volume - (EN)	Show details
	Using the QIAGEN® OneStep RT-PCR Kit with a 25 µl reaction volume and shorter reaction times for viral samples - (EN)	Show details
	Using the QIAGEN® OneStep RT-PCR Kit and Q-Solution® with a 25 µl reaction volume and shorter reaction times for viral samples - (EN)	Show details

Quick-Start Protocols (1)

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	QIAGEN OneStep RT-PCR Kit Quick-Start Protocol (EN)	Show details

Safety Data Sheets (4)

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	MSDS QIAGEN OneStep RT-PCR Kit (25)
	MSDS QIAGEN OneStep RT-PCR Kit (100)
	MSDS QIAGEN OneStep RT-PCR Kit (1000)
	CofA

References

Product citations in scientific articles

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品牌介绍

QIAGEN是一家专业化致力于生物分子样品制备解决方案的跨国经营企业，总部位于德国。1984年，QIAGEN在德国成立，1996年在美国纽约纳斯达克上市。QIAGEN拥有超过1000项专利和认可证明，在18个国家设立了分公司，代理商服务国超过40个，在全球有超过400000的用户。QIAGEN提供的产品超过500类，包括各种试剂，耗材和自动化纯化工作站。这些产品用于样品采集，稳定，核酸或蛋白的分离，纯化和检测中，不仅广泛的应用于科研领域的各个方面，在生物技术，制药，法医研究，食品安全检测，畜牧业和分子诊断领域也得到了广泛的应用。QIAGEN产品的卓越品质和在应用中的出色表现使得其成为样品处理中标准的代名词。

公司介绍

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发现与翻译研究人类识别与取证 Diagnostics & Clinical Research 仪器与自动化

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QIAGEN OneStep RT-PCR Kit

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Product Details

Wide range of annealing temperatures

Specifications

Product Resources

Product citations in scientific articles

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