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商品详细Qiagen/miScript PreAMP PCR试剂盒/用于12种预扩增反应：5x miScript PreAMP缓冲液、HOTSTAQ DNA聚合酶（2 U/µl）、miScript PreAMP通用引物、miR-16 miScript引物分析、SNORD95 miScript引物分析、miRNA逆转录酶联用

Qiagen/miScript PreAMP PCR试剂盒/用于12种预扩增反应：5x miScript PreAMP缓冲液、HOTSTAQ DNA聚合酶（2 U/µl）、miScript PreAMP通用引物、miR-16 miScript引物分析、SNORD95 miScript引物分析、miRNA逆转录酶联用

商品编号： 331451

品牌： Qiagen

市场价： ¥6520.00

美元价： 3912.00

产地：美国(厂家直采)

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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

Preamplification
Preamplification Kits2
REPLI-g10

miScript PreAMP PCR Kit

For preamplification of cDNA from samples containing low RNA amounts prior to real-time PCR analysis of miRNA

Breakthrough technology for miRNA expression analysis from limited RNA samples
Unbiased, reproducible results
Robust procedure
Fully integrated into the miScript PCR System workflow

The miScript PreAMP PCR Kit performs preamplification to enable use of the miScript PCR System for miRNA quantification with starting samples containing low RNA amounts. The miScript PreAMP PCR Kit uses highly multiplex, PCR-based preamplification of up to 400 miRNA-specific cDNA targets in one reaction. The miScript PreAMP PCR Kit, used with miScript PreAMP Primer Mixes, enables accurate and comprehensive expression analysis with as little as 10 ng total RNA. A single 10 ng cDNA synthesis reaction can be used as template for up to 10 preamplification reactions. This provides sufficient template for multiple Pathway-Focused miScript miRNA PCR Arrays or multiple replicates of a miRNome miScript miRNA PCR Array, depending on the miScript PreAMP Primer Mix used.

Important note: This product line has been discontinued and will be replaced by the miRCURY LNA miRNA PCR system. The remaining stock of kits is available for purchase through December 31, 2020. For more information about this transition, please visit our dedicated knowledge hub or contact us.

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Cat No./ID:331451

miScript PreAMP PCR Kit (12)

$326.00

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For 12 preamplification reactions: 5x miScript PreAMP Buffer, HotStarTaq DNA Polymerase (2 U/µl), miScript PreAMP Universal Primer, miR-16 miScript Primer Assay, SNORD95 miScript Primer Assay, miRNA reverse transcription control (miRTC), C. elegans miR-39 miScript Primer Assay, RNase-Free Water

Cat No./ID:331452

miScript PreAMP PCR Kit (60)

$1,463.00

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For 60 preamplification reactions: 5x miScript PreAMP Buffer, HotStarTaq DNA Polymerase (2 U/µl), miScript PreAMP Universal Primer, miR-16 miScript Primer Assay, SNORD95 miScript Primer Assay, miRNA reverse transcription control (miRTC), C. elegans miR-39 miScript Primer Assay, RNase-Free Water

The miScript PreAMP PCR Kit is intended for molecular biology applications. This product is not intended for the diagnosis, prevention, or treatment of a disease.

Product Details

High technical reproducibility.

cDNA was prepared from a human universal reference RNA sample using the miScript II RT Kit with miScript HiSpec Buffer. Three different users performed preamplification using the miScript PreAMP PCR Kit and miFinder miScript PreAMP Pathway Primer Mix. Preamplified cDNA was used for miRNA profiling in duplicate with the miFinder miScript miRNA PCR Array. High levels of correlation were observed between users when C_T values were [A] compared on a scatter plot or [B] plotted for all 3 users.

Extremely high correlation of expression between preamplified and nonpreamplified FFPE samples.

Preamplified cDNA and nonpreamplified cDNA from the same prep were used for miRNA profiling. Scatter plots of ΔC_T values (normalized against housekeeping controls) between normal and tumor sections demonstrate high correlation between nonpreamplified and preamplified samples. [A] 96-plex miFinder miScript PreAMP Pathway Primer Mix and Array or [B] 384-plex miScript PreAMP miRNome Primer Mix and Array were used. The miRNeasy FFPE Kit was used to purify RNA from normal and tumor lung tissue 5 µM FFPE sections. cDNA was prepared from 10 ng total RNA using the miScript II RT Kit with miScript HiSpec Buffer and preamplified using the miScript PreAMP PCR Kit.

Preamplification preserves expression patterns in FFPE samples using 1000-fold less input cDNA.

Preamplified cDNA and nonpreamplified cDNA from the same prep were used for miRNA profiling. Scatter plots of x-fold expression change calculations (2^-ΔΔCT) between normal and tumor sections demonstrate high correlation between nonpreamplified and preamplified samples. [A] 96-plex miFinder miScript PreAMP Pathway Primer Mix and Array or [B] 384-plex miScript PreAMP miRNome Primer Mix and Array were used. The miRNeasy FFPE Kit was used to purify RNA from normal and tumor lung tissue 5 µM FFPE sections. cDNA was prepared from 10 ng total RNA using the miScript II RT Kit with miScript HiSpec Buffer and preamplified using the miScript PreAMP PCR Kit.

100% increase in miRNAs detected from 10-fold less cDNA after preamplification from serum.

Total RNA was purified from 5 µl human serum using the miRNeasy Serum/Plasma Kit. cDNA was then prepared from 0.7 µl serum equivalence (SE) using the miScript II RT Kit with miScript HiSpec Buffer. cDNA (0.7 µl SE) was used directly for miRNA profiling or one-tenth of the cDNA preparation (0.07 µl SE) was preamplified using the miScript PreAMP PCR Kit with Serum & Plasma miScript PreAMP Pathway Primer Mix prior to profiling. miRNA profiling was performed with the with the Plasma & Serum miScript miRNA PCR Array. Plots of [A] mean C_T values achieved and [B] number of miRNAs detected demonstrate highly superior results from 10 fold less starting cDNA due to preamplification.

Performance

Highly significant increase in data from very limited sample amounts

The miScript PreAMP PCR Kit provides the opportunity to achieve large amounts of reliable miRNA profiling data from small samples and samples with very low RNA content. Previously unusable samples can now be used to elucidate valuable information about miRNA expression and function. Using preamplification, less than 1 µl serum equivalence RNA can be used to robustly profile miRNA expression using the Serum & Plasma miScript miRNA PCR Array (see figure "100% increase in miRNAs detected from 10-fold less cDNA after preamplification from serum").

High correlation with results from nonpreamplified samples

Reliable, unbiased amplification using the miScript PreAMP PCR Kit provides data that correlates highly with nonpreamplified sample, ensuring high confidence in results. Comparison of miRNA profiling results from tumor and normal lung tissue FFPE samples demonstrated this strong correlation (see figures "Preamplification preserves expression patterns in FFPE samples using 1000 fold less input cDNA" and "Extremely high correlation of expression between preamplified and nonpreamplified FFPE samples").

Reproducible results and robust procedure

Preamplification using the miScript PreAMP PCR Kit is a robust procedure that ensures consistent, reproducible results. Highly overlapping results were achieved when preamplification and miRNA profiling experiments were performed by different users (see figure "High technical reproducibility").

Principle

The miScript PCR System enables sensitive, specific miRNA quantification and profiling using SYBR^® Green real-time PCR. The miScript PCR System covers all the steps involved in conversion of RNA to cDNA and subsequent real-time PCR detection of miRNAs. The miScript PreAMP PCR Kit is a component of the miScript PCR System that allows researchers to perform miRNA profiling experiments using very limited amounts of starting RNA (see Janas, M.M. et al. (2012) Reduced Expression of Ribosomal Proteins Relieves MicroRNA-Mediated Repression. Molecular Cell 46, 171.). This is necessary when working with samples that contain low amounts of RNA, such as body fluids, formalin-fixed, paraffin-embedded (FFPE) samples, and low cell number samples, such as laser capture microdissection (LCM) samples, flow-sorted cells, circulating tumor cells, and fine-needle biopsies. The low RNA yields obtained from small samples are often insufficient for reliable miRNA profiling experiments, even when using sensitive techniques such as real-time RT-PCR.The miScript PreAMP PCR Kit, used with miScript PreAMP Primer Mixes, is a breakthrough technology enabling accurate and comprehensive expression analysis with as little as 10 ng total RNA. A single 10 ng cDNA synthesis reaction can be used as template for up to 10 preamplification reactions. This provides sufficient template for multiple Pathway-Focused miScript miRNA PCR Arrays or multiple replicates of a miRNome miScript miRNA PCR Array, depending on the miScript PreAMP Primer Mix used. The miScript PreAMP PCR Kit uses highly multiplex, PCR-based preamplification of up to 400 miRNA-specific cDNA targets in one reaction. Typically, preamplification results in a 2000–4000-fold amplification in either a 96-plex or 384-plex reaction.

Procedure

miRNA profiling from low-quantity RNA samples requires the following 3 steps: reverse transcription using the miScript II RT Kit, preamplification using the miScript PreAMP PCR Kit and miScript PreAMP Primer Mix, and real-time PCR using the corresponding miScript miRNA PCR Array and the miScript SYBR Green PCR Kit. Preamplification using the miScript PreAMP PCR Kit and miScript PreAMP Primer Mix is therefore fully integrated into the miScript PCR System miRNA profiling workflow, ensuring robust protocols and reliable, reproducible results.

miScript PreAMP Primer Mixes are available for all miScript miRNA PCR Arrays. Alternatively, if desired, preamplification can be performed using the miScript PreAMP PCR Kit together with a mix of up to 25 miScript Primer Assays.

Preamplification controls

The miScript PreAMP PCR Kit includes 4 miScript Primer Assays for use in preamplification control experiments to test preamplified cDNA. Controls and their purpose are listed below. For more detailed information, see the miScript PreAMP Handbook.


Control	Purpose
miR-16 miScript Primer Assay	Determination of the optimal dilution factor for preamplified cDNA if the template starting amount is unknown
SNORD95 miScript Primer Assay	Data normalization for tissue and cell samples using the ∆∆CT method of relative quantification Determination of the optimal dilution factor for preamplified cDNA if the template starting amount is unknown and miR-16 is not present
C. elegans miR-39 miScript Primer Assay	Measurement of miRNeasy Serum/Plasma Spike-In Control (if added during purification) for determination of recovery from serum and plasma samples
miRTC miScript Primer Assay	Assessment of reverse-transcription efficiency

Applications

The miScript PreAMP PCR Kit is used as part of the miScript PCR System for:

Mature miRNA quantification and profiling

Biomarker discovery from serum or plasma

Product Resources

Application Notes (1)

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	(EN) - Serum miRNA and lncRNA detection as a potential biomarker of lung cancer	Show details

Brochures & Guides (1)

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	Total RNA Discovery - (EN) Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow	Show details

Kit Handbooks (2)

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		Show details
	(EN) - miScript PreAMP Handbook For preamplification of cDNA from samples containing low RNA amounts prior to real-time PCR analysis of microRNA	Show details

Quick-Start Protocols (1)

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	Quick-Start Protocol miScript PreAMP PCR Kit and miScript PreAMP Primer Mixes - (EN)	Show details

White Papers (1)

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	miRNA biomarker discovery - overcoming limiting sample material	Show details

Safety Data Sheets (2)

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	MSDS miScript PreAMP PCR Kit (12)
	MSDS miScript PreAMP PCR Kit (60)

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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

品牌分类

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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miScript PreAMP PCR Kit

Buy Products

Product Details

Highly significant increase in data from very limited sample amounts

High correlation with results from nonpreamplified samples

Reproducible results and robust procedure

Preamplification controls

Product Resources

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