qPCR ProbesMaster with ROX | Sondas / Detecção TaqMan® - Cellco
Master Mix for real time PCR using labeled DNA probes with ROX

For in vitro use only!

Envio:
Shipped on blue ice

Condições de armazenamento:
Store at -20 °C
(Avoid freeze/thaw cycles, store in dark). Store at 4 °C for up to 3 months possible.

Validade:
12 months

Forma:
Liquid

Kit contents:
qPCR ProbesMaster (#PCK-110 - red cap)
qPCR Pol, dATP, dCTP, dGTP, dUTP, reaction buffer with KCl, (NH₄)₂SO₄, MgCl₂ and stabilizers

ROX reference dye (#PCK-121)
25 µM ROX Reference Dye.

Dual-labeled DNA Probes (NOT PROVIDED)
Real-time PCR technology based on dual-labeled DNA probes provides a high sensitive and high specific PCR system with multiplexing capability. It requires two standard PCR primers and the DNA probe that hybridizes to an internal part of the amplicon. The sequence of the dual-labeled DNA probe should avoid secondary structure and primer-dimer formation.

Description:
qPCR ProbesMaster + ROX is designed for the quantitative realtime analysis of DNA samples using DNA probe based detection. The master mix is recommended for use with Dual Labeled Fluorescent Probes, e.g. TaqMan®, Molecular Beacons or FRET probes. It provides an easy-to-handle and powerful tool for quantification of sample DNA in a broad dynamic range of up to 6 orders of magnitude with exceptional sensitivity and precision. The Master contains all reagents required for qPCR (except template, primer and labeled fluorescent probe) in a premixed 2x concentrated ready-to-use solution. The high specificity and sensitivity of the mix is achieved by an optimized hot-start polymerase. Its activity is blocked at ambient temperature and switched on automatically at the onset of the initial denaturation. The thermal activation prevents the extension of nonspecifically annealed primers and primer-dimer formations at low temperatures during PCR setup. The mix contains dUTP instead of dTTP and allows an UDG (Uracil-DNA-Glycosylase) treatment at the onset of thermal cycling to prevent carry-over contaminations of DNA from previous PCR reactions. The reaction chemistry of the kit is optimized for block-based PCR instruments that are compatible with the evaluation of the ROX reference signal.

Preparation of the qPCR master mix:
The preparation of a master mix is crucial in quantitative PCR reactions to reduce pipetting errors. Prepare a master mix of all components except template as specified. A reaction volume of 20-50 µl is recommended for most real-time instruments. Prepare 13 volumes of master mix for 12 samples or a triple-set of 4 samples. Pipet with sterile filter tips and minimize the exposure of the labeled DNA probe to light. Perform the setup in an area separate from DNA preparation or analysis. No-template controls should be included in all amplifications.