technique(s)
cDNA synthesis: suitable
shipped in
wet ice
storage temp.
−20°C
General description
MystiCq ®microRNA cDNA Synthesis Mix 试剂盒从总 RNA 或预富集样品或 microRNA 开始，包含将成熟 microRNA 转化为 qPCR 的 cDNA 模板所需的所有成分。 由于 microRNA 不是天然进行多聚腺苷酸化，因此 cDNA 合成过程的第一步是通过 poly(A) 聚合酶反应使 microRNA多聚腺苷酸化。 接下来，添加 ReadyScript 逆转录酶、oligo-dT 接头引物和其它必要试剂，将多聚腺苷酸化的 microRNA 转化为 cDNA。 接头引物的5' 端包含一个与 MystiCq 通用 PCR 引物的序列互补的独特序列，该序列是MystiCq microRNA qPCR 检测系统的另一个成分。该试剂盒还包含一个针对 SNORD44 的特异性阳性对照引物， SNORD44 是一种在大多数人体组织中广泛表达的小核仁 RNA。 试剂盒中包括足量的Poly(A)拖尾缓冲液和 MicroRNA cDNA 反应混合物，以允许不使用 poly(A) 聚合酶和不使用逆转录酶的对照反应。为了分析单个 microRNA，将 MystiCq 通用 PCR 引物（靶向接头引物序列）与 MystiCq® microRNA qPCR Assay 引物和 MystiCq microRNA SYBR ™Green qPCR ReadyMix ™一起使用。
Application
MystiCq® 微小 RNA cDNA 合成混合物已用于通过逆转录 miRNA 第一链 cDNA 合成，用于从总 RNA 开始的定量聚合酶链式反应（qPCR）
Features and Benefits
- 如果需要，单独的多腺苷酸化和逆转录步骤允许优化方案的可能性
- 生成的 cDNA 可存档备用
- 单一 RT 反应可提供多达 1000 个 qPCR 反应
- 包括用于大多数人体组织的阳性对照引物
Packaging
默认反应体积为 20μL
Legal Information
SYBR is a trademark of Thermo Fisher Scientific or its subsidiaries
MystiCq is a registered trademark of QIAGEN Beverly Inc.
ReadyMix is a trademark of Sigma-Aldrich Co. LLC
仅试剂盒组分
产品编号
说明
- Poly (A) Tailing Buffer (5X)
- Human Positive Control Primer
- Nuclease-free Water
- MystiCq® Universal PCR Primer
- Poly (A) Polymerase
- microRNA cDNA Reaction Mix
- ReadyScript® Reverse Transcriptase
存储类别
10 - Combustible liquids
法规信息
常规特殊物品
此项目有
实验方案
Method for purification, reverse transcription and quantitative PCR for MicroRNAs using Mysticq reagents
商品
Sigma’s Imprint RNA Immunoprecipitation Kit was used to copurify human argonaute 2 (Ago2)-associated RNAs from HeLa cells. MicroRNAs reverse transcribed and quantitating using Mysticq reagents.
PCR master mix simplifies PCR/RT-PCR with components like DNA polymerase, dNTPs, MgCl2, and buffer, available commercially or DIY.
Real-time polymerase chain reaction allows researchers to estimate the quantity of starting material in a sample. It has a much wider dynamic range of analysis than conventional PCR
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Instructions
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