Description

Description: Direct qPCR ProbesMaster highROX is designed for quantitative real-time analysis of target DNA directly from blood, swabs and animal- or plant tissue. The mix allows robust amplification avoiding the requirement of any prior DNA purification procedures. The mix is recommended for use with dual-labeled fluorescent probes, e.g. TaqMan®, Molecular Beacon or Scorpion probes. It provides a powerful tool for multiplex-quantification of sample DNA in a broad dynamic range with exceptional sensitivity and precision. The mix contains all reagents required for qPCR (except template, primer and labeled fluorescent probe) in a premixed 2 x concentrated ready-to-use solution. High robustness, reliability and sensitivity of the mix are based on a an antibody-blocked hot start polymerase in combination with an optimized and well-balanced buffer system. The mix ensures fast and easy preparation with a minimum of pipetting steps and is specially recommended for: Direct detection of viral or bacterial DNA in nasal or throat swabsDirect PCR from blood samplesDirect amplification of target DNA from various tissue samplesPoint-of-Care diagnostics. Multiplexing Capability Real-time PCR technology based on dual-labeled DNA probes provides a high sensitive and specific PCR system with multiplexing capability. The simultaneous detection of multiple targets in a single tube requires a primer/probe set for each target amplification. Sequences and concentrations of primers and probes should be optimized to avoid mutual influence, secondary structures and primer-dimer formations. Amplification of each target is detected in a separate fluorescence channel. Interference of remaining components from sample matrix Due to the fast and easy but relatively rough sample preparation remaining components from the sample matrix may be co-transferred into the PCR assay. These remains are mostly blocked by a combination of specially optimized additives. If inhibition of the PCR reaction occurs with higher volumes of transferred sample volume, please reduce the sample volumes or use a dilution of the sample in 1 x Direct Extraction Buffer or water. Remaining components of the sample matrix may also show fluorescence signals specially in the yellow and red spectral range. If using this fluorescence range for multiplex PCR assays or if using ROX, please take special attention that there is no interference between fluorescence from remaining matrix material and the used fluorescence channels for amplicon detection. ROX reference dye Direct qPCR ProbesMaster highROX contains 500 nM ROX passive reference dye in the final assay. The dye does not take part in the PCR reaction but allows to normalize for non-PCR related signal variation and provides a baseline in multiplex reactions. The reaction chemistry of the kit is optimized for instruments that are compatible with the evaluation of a high ROX reference signal.

Additional Text

Concentration

2 X

Storage Note

store at -20 °C|12 months|avoid freeze/thaw cycles,|stable at 4 °C for up to 4 weeks