Phusion^® RT-PCR Kit

A complete two-step kit for RT-PCR

Note: Starting May 2010 the Phusion Hot Start DNA polymerase in the kit will be replaced with the new Phusion Hot Start II DNA Polymerase. The new polymerase allows more flexibility to primer design. No changes are needed to excisting protocols or primers.

Description

Finnzymes’ Phusion® RT-PCR Kit is a complete two-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 in PCR. The superior features of Phusion Hot Start II DNA Polymerase enable 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.

General information about Phusion DNA Polymerases, their advantages and applications can be found on Phusion DNA Polymerase products page.

Applications

• Detection of gene expression
• Generation of cDNA products with superior fidelity for cloning and sequencing

Advantages

• High yields for a broad range of RT-PCR products
• Accurate amplification of cDNA with Phusion Hot Start II DNA Polymerase (52x Taq fidelity)
• Short and simple cDNA synthesis and PCR protocols
• Easy to use; the kit includes all necessary components for performing cDNA synthesis and PCR

Phusion® RT-PCR system is robust and amplifies long fragments

Click pictures for a full-sized image
Robustness of Phusion RT-PCR Kit	Amplification of long fragments

Components

The kit contains sufficient materials for 100 (F-546L) or 20 reactions (F-546S) including control reactions.

Phusion® RT-PCR Kit
Component	Concentration	F-546S	F-546L
RT enzyme mix (M-MuLV RNase H+ + RNase inhibitor)		1 x 40 µl	1 x 200 µl
10x RT Buffer (includes 50 mM MgCl2*)		1 x 1.5 ml	1 x 1.5 ml
Oligo(dT)15 primer	100 ng/µl	1 x 20 µl	1 x 100 µl
Random primers (hexamers)	50 ng/µl	1 x 20 µl	1 x 100 µl
dNTP mix	10 mM	1 x 40 µl	2 x 100 µl
Phusion® Hot Start II DNA Polymerase	2 U/µl	1 x 10 µl	1 x 50 µl
5x Phusion® HF Buffer		1 x 1.5 ml	1 x 1.5 ml
Control RNA with carrier	10 ng MS2 RNA/µl 30 µg E. coli ribosomal RNA/ml	1 x 20 µl	1 x 20 µl
Control Primer Mix	25 µM each	1 x 20 µl	1 x 20 µl

* Provides 5 mM MgCl₂ in 1x reaction concentration.

Storage and stability

Store all kit components at -20 °C. All the kit components can be refrozen and stored at -20°C without affecting the performance of the kit. The Phusion RT-PCR Kit is stable for one year from the date of packaging when stored and handled properly.

Ordering info

Phusion® RT-PCR Kit
F-546S	20 reactions
F-546L	100 reactions

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