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Qiagen/miScript PreAMP PCR Kit/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 con
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PreamplificationPreamplification Kits2REPLI-g10miScript PreAMP PCR KitPrint 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 samplesUnbiased, reproducible resultsRobust procedureFully integrated into the miScript PCR System workflowThe 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.Buy ProductsProduct DetailsProduct ResourcesBuy ProductsCat No./ID:331451miScript PreAMP PCR Kit (12)$326.00Order ProductFor 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 WaterCat No./ID:331452miScript PreAMP PCR Kit (60)$1,463.00Order ProductFor 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 WaterThe 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 CT 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 ΔCT 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 CT 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"). PrincipleThe 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.ProceduremiRNA 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 plasmaProduct ResourcesYou are not authorized to download the resourceApplication Notes (1) Sort optionsSort alphabetically (EN) - Serum miRNA and lncRNA detection as a potential biomarker of lung cancerShow detailsShow allShow lessBrochures & Guides (1) Sort optionsSort alphabetically Total RNA Discovery - (EN)Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflowShow detailsShow allShow lessKit Handbooks (2) Sort optionsSort alphabetically Show details(EN) - miScript PreAMP HandbookFor preamplification of cDNA from samples containing low RNA amounts prior to real-time PCR analysis of microRNAShow detailsShow allShow lessQuick-Start Protocols (1) Sort optionsSort alphabetically Quick-Start Protocol miScript PreAMP PCR Kit and miScript PreAMP Primer Mixes - (EN)Show detailsShow allShow lessWhite Papers (1) Sort optionsSort alphabetically miRNA biomarker discovery - overcoming limiting sample materialShow detailsShow allShow lessSafety Data Sheets (2) Sort optionsSort alphabetically MSDS miScript PreAMP PCR Kit (12)MSDS miScript PreAMP PCR Kit (60)Show allShow lessfragment fix placeholderCustomers who bought these products also boughtCat No./ID:331242miScript PreAMP Pathway HC Primer Mix60 µl primer mix for 12 preamplification reactions; for use with a miScript miRNA HC PCR ArraySelect TargetsCat No./ID:218160miScript II RT Kit (12)For 12 cDNA synthesis reactions: miScript Reverse Transcriptase Mix, 10x miScript Nucleics Mix, 5x miScript HiSpec Buffer, 5x miScript HiFlex Buffer, RNase-Free Water$146.00Add To CartCat No./ID:218073miScript SYBR Green PCR Kit (200)For 200 reactions: QuantiTect SYBR Green PCR Master Mix, miScript Universal Primer$494.00Add To CartCat No./ID:218075miScript SYBR Green PCR Kit (1000)For 1000 reactions: QuantiTect SYBR Green PCR Master Mix, miScript Universal Primer$2,135.00Add To CartCat No./ID:218161miScript II RT Kit (50)For 50 cDNA synthesis reactions: miScript Reverse Transcriptase Mix, 10x miScript Nucleics Mix, 5x miScript HiSpec Buffer, 5x miScript HiFlex Buffer, RNase-Free Water$539.00Add To CartCat No./ID:218076miScript SYBR Green PCR Kit (2000)For 2000 reactions: QuantiTect SYBR Green PCR Master Mix, miScript Universal Primer$3,075.00Add To CartCat No./ID:180492QIAseq Ultralow Input Library Kit (12)For 12 reactions: buffers and reagents for Ultralow Input End Polishing, Ultralow Input Ligation and HiFi library amplification. 12-plex adapters sold separately. For use with Illumina instruments.$342.00Add To CartCat No./ID:217184miRNeasy Serum/Plasma Kit (50)For 50 total RNA preps: 50 RNeasy MinElute Spin Columns, Collection Tubes (1.5 ml and 2 ml), QIAzol Lysis Reagent, Ce_miR-39_1 miScript Primer Assay, RNase-free Reagents and Buffers$504.00Add To CartCat No./ID:330451RT2 PreAMP cDNA Synthesis Kit (12)RT2 Nano PreAMP cDNA Synthesis Kit$564.00Add To CartCat No./ID:1069275Buffer ACB (300 ml)Buffer ACB (300 ml)$174.00Add To CartCat No./ID:292015mericon Pig Kit (96)For 96 reactions: PCR Assay Pig, Internal Control, Positive Control, Multiplex PCR Master Mix, QuantiTect Nucleic Acid Dilution Buffer, RNase-free water$1,087.00Add To CartCat No./ID:330521RT² SYBR Green ROX qPCR Mastermix (8)Contains 8 x 1.35 ml tubes: for 8 x 96-well RT² PCR Arrays, 4 x 384-well RT² PCR Arrays or 800 x 25 µl reactionsor 2000 x 10µlreactions$818.00Add To CartCat No./ID:330501RT² SYBR Green qPCR Mastermix (8)Contains 8 x 1.35 ml tubes: for 8 x 96-well RT² PCR Arrays, 4 x 384-well RT² PCR Arrays or 800 x 25 µl reactionsor 2000 x 10µlreactions$804.00Add To CartCat No./ID:218300miScript Primer Assay (100)10x miScript Primer Assay (contains one miRNA-specific primer)Select TargetsCat No./ID:76603miRCURY Exosome Serum/Plasma KitFor isolation of exosomes and other extracellular vesicles from serum or plasma; includes 10 ml Precipitation Buffer A, 10 ml Resuspension Buffer, 200 U Thrombin and 500 µl Thrombin Buffer$345.00Add To CartCat No./ID:59824EpiTect Fast DNA Bisulfite Kit (50)For 50 preps: Bisulfite Solution, DNA Protect Buffer, MinElute DNA Spin Columns, Carrier RNA, and Buffers$236.00Add To CartCat No./ID:37502Qproteome Cell Compartment KitFor up to 10 subcellular fractionations*: Extraction buffers, Protease Inhibitor Solution, Benzonase®.$332.00Add To CartCat No./ID:204363QuantiTect Probe PCR +UNG Kit (200)For 200 x 50 µl reactions: 3 x 1.7 ml 2x QuantiTect Probe PCR Master Mix, 100 ul UNG Solution, 2 x 2 ml RNase-Free Water$598.00Add To CartCat No./ID:30410Ni-NTA Superflow (25 ml)25 ml nickel-charged resin (max. pressure: 140 psi)$421.00Add To CartCat No./ID:158489Gentra Puregene Blood Kit Plus (1000 ml)For 1000 ml blood: RBC Lysis Solution, RNase A Solution, and Reagents$1,687.00Add To Cart

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。


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