Titan One Tube RT-PCR System / Kit

for one-step RT-PCR amplification of up to 6 kb targets Cat. No. 1 888 382      25 reactions

Cat. No. 1 855 476    100 reactions
Cat. No. 1 939 823      50 reactions

Description

Components of the Titan One Tube RT-PCR System/Kit allow completion of RT-PCR in a single tube, one-step reaction. The system includes the following components:

Enzyme mix containing AMV Reverse Transcriptase (for reverse transcription) and the Expand High Fidelity PCR System (Taq/Pwo enzyme blend, for PCR)
Additional reaction components (RT-PCR reaction buffer with 7.5 mM MgCl₂; MgCl₂ solution; dithiothreitol solution) for convenient, quick reaction set-up

In addition to all the components of the Titan One Tube RT-PCR System, the Titan One Tube RT-PCR Kit also contains:

PCR Grade dNTP mix
PCR grade water
RNase Inhibitor; human control RNA
Control primer mix for amplification of a human beta-actin product

Application

The Titan One Tube RT-PCR System or Kit can generate up to 6 kb PCR products from total RNA, mRNA, or viral RNA. The system may be used for:

Amplification of specific RNA targets
Cloning of RNA-sequences without constructing a cDNA library
Competitive or multiplex PCR.

Application profile

Operating parameters

Starting samples: 1 pg to 1 µg total RNA
Temperature optimum, reverse transcriptase reaction: 50°C
Note: The enzyme may be used at temperatures up to 60°C
Requires two sequence-specific primers
Accepts DIG-labeled and other modified nucleotides

Note: The Titan One Tube RT-PCR System/Kit is not recommended for carry-over prevention using dUTP and Uracil-DNA Glycosylase

Experimental result

RT-PCR of beta-actin transcript from K562 cells. mRNA from serially diluted cell lysates was captured in streptavidin-coated PCR tubes using the mRNA Capture Kit. RT-PCR was performed with the Titan System (RT at 50°C) or RNase H^- M-MuLV reverse transcriptase fromSupplier L (RT at 42°C) +Taq DNA polymerase according to each supplier’s protocol. The TitanSystem delivers > 100 times greater sensitivity than the two step methodwith Supplier L.

Key advantages

Significantly reduces time needed to perform RT-PCR, because the one-step reaction minimizes handling and reduces the risk of contamination
Improves the specificity and sensitivity of cDNA synthesis, because the elevated temperature of the AMV Reverse Transcriptase reaction eliminates secondary structure in the template and increases primer specificity
Increases yield and fidelity of PCR amplification, because the Taq/Pwo enzyme blend gives threefold greater fidelity and twofold higher yields than Taq DNA Polymerase alone

Related Products

Roche Contact Information

Roche

Roche Diagnostics Corporation
P.O. Box 50414
9115 Hague Road
Indianapolis, IN 46250-0414 USA

Customer Service: 800-262-1640

Tech Support: 800-262-4911

Fax Number: 800-428-2883

Web Site: http://www.roche-applied-science.com

Titan One Tube RT-PCR System / Kit