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Protocols for Phusion™ RT-PCR Kit FAQs for Polymerases Technical Reference for Polymerases Enzyme Finder NEBcutter NEBuffer Chart Double Digest Finder Isoschizomers DNA Sequences and Maps REBASE Phusion™ HF Buffer Pack Phusion™ Hot Start High-Fidelity DNA Polymerase

Home > Products > Polymerases > RT-PCR and qRT-PCR > Phusion™ RT-PCR Kit Phusion™ RT-PCR Kit New | Developed & Manufactured By Finnzymes Oy. Distributed by New England Biolabs, Inc. Catalog # Size Concentration Price Qty   F-546L 100 reactions   $380.00 F-546S 20 reactions   $95.00 Prices are in US dollars and valid only for US orders. Download: MSDS PDF A complete two-step kit for RT-PCR 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 to perform 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 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 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. Robustness of different RT-PCR systems. A typical RT-PCR experiment for amplifying short cDNA fragments (192-1113 bp). Finnzymes' RT-PCR system was compared to the RT-PCR systems from two major suppliers. Total RNA from human skeletal muscle was transcribed with random priming according to each supplier's recommendations. Amplification of cDNA was performed with a high-fidelity DNA polymerase recommended by the suppliers. Due to the various sizes of amplicons, a PCR protocol with the lowest annealing temperature and the longest extension time defined by amplicons was used. Robust Phusion RT-PCR Kit produced all 11 amplicons with highest yields. With competitors' RT-PCR systems, less amplicons with varying yields were transcribed and amplified. M is a molecular size marker. Phusion RT-PCR Kit is able to amplify long RT-PCR fragments. 3.3 kb, 6 kb, 8 kb and 10 kb fragments of dystrophin gene were reverse transcribed and amplified with Phusion RT-PCR Kit. Oligo (dT) was used for priming. Starting template was human skeletal muscle total RNA (50 ng). M is a molecular size marker. Applications: Detection of gene expression Generation of cDNA products with superior fidelity for cloning and sequencing Advantages: Robust: broad range of RT-PCR products with high yields Accurate: cDNA amplification with 52x Taq fidelity Fast: short and simple cDNA synthesis and PCR protocols Easy to use: complete 2-step kit for RT-PCR Kit Components: Control Primer Mix (25 μM each) Control RNA with carrier dNTP Mix (10 mM) Oligo(dT)15 primer (100 ng/μl) Phusion™ HF Buffer Pack (5X) Phusion™ Hot Start High-Fidelity DNA Polymerase (2 units/μl) Random Hexamers (50 ng/μl) RT enzyme mix (M-MuLV RNase H+ + RNase inhibitor) RT Reaction Buffer (10X) Storage Conditions Storage Temperature: -20°C Notes General notes: 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. Reagents Sold Separately Phusion™ HF Buffer Pack Phusion™ Hot Start High-Fidelity DNA Polymerase