RT2 First Strand Kit

用于更快、更高效地合成cDNA,并去除RNA样本中的基因组DNA

Products

RT2 First Strand Kit 旨在用于分子生物学应用。该产品不能用于疾病诊断、预防和治疗。
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RT2 First Strand Kit (12)

Cat. No. / ID:  330401

For twelve 20 µl first strand cDNA synthesis reactions; Buffer GE (24 µl), 5x Buffer BC3 (48 µl), RE3 Reverse Transcriptase Mix (24 µl), Control P2 (12 µl), Nuclease-Free Water (1 ml)
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RT2 First Strand Kit (50)

Cat. No. / ID:  330404

For fifty 20 µl first strand cDNA synthesis reactions; Buffer GE (100 µl), 5x Buffer BC3 (200 µl), RE3 Reverse Transcriptase Mix (100 µl), Control P2 (50 µl), Nuclease-Free Water (1 ml)
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Features

  • 快速、高效地合成cDNA
  • 完全去除基因组DNA
  • 每次反应只需使用25 ng总RNA,即可获得结果
  • 获得的cDNA可直接用于real-time PCR
  • RNA外参可监测酶抑制剂

Product Details

RT2 First Strand Kit可通过快速、方便的操作流程高效合成cDNA,并去除RNA样本中的基因组DNA。合成的cDNA可即用于多基因real-time PCR表达分析。

Principle

RT2 First Strand Kit可通过快速、方便的操作流程从mRNA高效合成cDNA,用于基因表达分析。

该试剂盒专有的操作流程,可在逆转录前有效去除RNA样本中的基因组DNA。使用随机引物和oligo-dT引物可进行无偏向性的逆转录,逆转录合成的cDNA产量高和长度合适。内置式RNA外参可帮助监测逆转录效率并检测酶抑制剂。RT2 First Strand Kits非常适合基于real-time PCR的基因表达分析,可配合 RT2 Profiler PCR Arrays、RT2 qPCR Primer Assays和其他商业化基因表达分析使用。

Procedure

专有的Buffer GE与样本混合后在热循环仪上孵育。加入酶和另外的缓冲液后,样本可在多种热循环仪上逆转录。转录的cDNA可直接用于下游应用。推荐使用RT² Profiler PCR Arrays或RT² qPCR Primer进行进一步分析。

 

RT² First Strand Kits每次反应使用至少25 ng或至多5 µg总RNA都可获得结果。但是,最佳起始样本量取决于目标转录本的相对丰度。低丰度转录本需要较多RNA;高丰度转录本需要较少RNA。在此方法的线性范围内,更大起始量的RNA可获得更多的阳性信号(如基因表达检测)。

 

Applications

使用RT2 First Strand Kit合成的cDNA可在多基因real-time PCR表达分析中获得出色的结果。

Resources

安全数据表 (1)
Download Safety Data Sheets for QIAGEN product components.
试剂盒操作手册 (3)
For pathway-focused gene expression profiling using real-time RT-PCR
产品介绍与指南 (1)
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
仪器技术参数 (1)
For pathway-focused gene expression analysis
Certificates of Analysis (1)
Brochures & Guides (1)
Simultaneously profile mRNA, miRNA and lncRNA using a simple, complete workflow
Instrument Technical Documents (1)
For pathway-focused gene expression analysis
Kit Handbooks (3)
For pathway-focused gene expression profiling using real-time RT-PCR

FAQ

What do I need to complete a RT² qPCR Primer Assay?

You need:

  1. A RT² SYBR Green Mastermix that matches the qPCR instrument in your laboratory;
  2. RT² qPCR Primer Assays for your target genes;
  3. A Housekeeping gene RT² qPCR Primer Assay.

We also recommend using our RT² First Strand Kit for reverse transcription.

FAQ ID -2707
What is the recommended solution in which to store RNA samples that will be used as templates for cDNA synthesis?
For best results, all RNA samples should be suspended in RNase-free water. Alternatively, RNase-free 1 mM sodium citrate (pH 6.5) or 10 mM Tris buffer (pH 7.0) may be used. Do not use DEPC-treated water, as most DEPC preparations are contaminated with molecules that are inhibitory to reverse transcription and/or PCR. For long-term storage, RNA preps may be stored at -70 ºC in RNase-free water, or the buffers listed above, or precipitated in ethanol or isopropanol. In order to avoid repeated freeze-thaw cycles, it is recommended that frozen RNA samples be stored as multiple, single-use aliquots.
FAQ ID -2659
How can I avoid or remove genomic DNA contamination from the total RNA preparation?

Carry out all procedures in a "DNA-free" workspace (see FAQ 2654). Be sure to include any DNase treatment steps in the recommended RNA isolation procedure or treat RNA separately with RNase-free DNase followed by repurification using a spin-column based method. RNeasy Mini Kit can be easily combined with RNase-free DNase. Alternatively, kits like the RNeasy Plus Universal Tissue already include a DNA removal step. Be sure to double both the units of enzyme and the incubation time recommended by the RNase-free DNase manufacturer. To minimize DNA contamination in your RNA preparations, and avoid the need for supplemental DNase treatments, we recommend using the RT2 First Strand Kit, which includes a highly efficient genomic DNA elimination step before reverse transcription.

 

NOTE: Our Chemistries are not compatible with AMBION's Turbo DNA-Free Kits.

FAQ ID -2662