Phusion RT-PCR Kit

• 产品编号：F-546S      品牌：Finnzymes       原厂货号：F-546S
• 产品分类：PCR/RT-PCR/qRT-PCR > RT-PCR/RT-qPCR > RT-PCR（Endpoint）
• 应用分类：

其它组分：

描述：

Thermo Scientific Phusion RT-PCR Kit用于两步法高保真RT-PCR实验。该试剂盒包括cDNA合成和PCR扩增所需的全部试剂。第一步中，M-MuLV 逆转录酶可以以不同种类的RNA为模板，并利用各类引物高效合成cDNA。在第二步中，Phusion Hot Start II High Fidelity DNA Polymerase则可用于cDNA的PCR扩增。这种聚合酶有着优异的保真度和扩增能力，且带有热启动修饰，可以使PCR反应更为特异。Phusion Hot StartII High Fidelity DNA Polymerase的这些优异特性，不仅可以使cDNA扩增的保真度更高，还能在缩短循环时间的同时，提高扩增产物的得率。当需要制备用于克隆和基因表达研究的cDNA时，Phusion RT-PCR试剂盒是一种理想的选择。

优点：
• 产量高
• cDNA扩增保真度高，其保真度是Taq酶的52倍
• 简单快速的cDNA合成和PCR扩增
• 易用：完整的两步法RT-PCR试剂盒

应用：
• 基因表达分析
• 制备具有优异保真度的cDNA产物，以用于克隆和测序


使用Phusion RT-PCR系统进行强劲而高产的cDNA扩增
在扩增短cDNA片段(192-1113bp)的典型RT-PCR实验中，将Phusion RT-PCR 系统与市场上两个主要供应商的RT-PCR系统进行了比较。用随机引物转录来自人骨骼肌的总RNA。用厂家各自推荐的高保真DNA聚合酶进行cDNA扩增。由于扩增子长度不同，因此在PCR反应中，采用的是最低的退火温度和最长的延伸时间。强劲的Phusion RT-PCR Kit扩增得到了全部11个扩增子，且保持了最高的产物得率。


Phusion RT-PCR Kit扩增长RT-PCR片段
使用Phusion RT-PCR Kit 对长度为3.3kb、6kb、8kb和10kb的人dystrophin mRNA进行了逆转录和扩增。使用了Oligo (dT)16引物，起始模板为人骨骼肌总RNA(50 ng) 。

原厂资料：

Thermo Scientific Phusion RT-PCR Kit is a complete 2-step kit designed for high-fidelity RT-PCR. The kit contains the full set of reagents required for performing cDNA synthesis and PCR in two steps.

In the first step, a variety of RNA templates and all priming options may be used for efficient synthesis of cDNA with M-MuLV Reverse Transcriptase. In the second step, Phusion Hot Start II High-Fidelity DNA Polymerase is used for cDNA amplification. This polymerase is extremely accurate and robust, and the hot start modification guarantees additional specificity during PCR, enabling accurate cDNA amplification with high yields and short cycling times. Phusion RT-PCR Kit is an ideal choice when producing cDNA for cloning and gene expression studies.

Highlights
• Broad range of RT-PCR products with high yields
• Accurate cDNA amplification with 52x Taq fidelity
• Short and simple cDNA synthesis and PCR protocols
• Easy to use: complete 2-step kit for RT-PCR

Applications
• Gene expression analysis
• Generation of cDNA products with superior fidelity for cloning and sequencing

Includes
• RT enzyme mix (M-MuLV RNase H+ + RNase inhibitor)
• 10x RT Buffer (includes 50 mM MgCl2)
• Oligo(dT)15 primer (100 ng/μL)
• Random primers (hexamers, 50 ng/μL)
• 10 mM dNTP mix
•Phusion Hot Start II DNA Polymerase (2 U/μL)
• 5x Phusion HF Buffer
• Control RNA with carrier (10 ng MS2 RNA/µL, 30 µg E. coli ribosomal RNA/mL)
• Control Primer Mix (25 μM each)