qPCR MultiplexMaster highROX, Master mix for multiplex real-time PCR

Catalogue Number: PCR-323S-JEN

Manufacturer:Jena Bioscience
Shelf Life:12 months
Physical state:Liquid
Type:qPCR Mixes
Shipping Condition:Blue Ice
Unit(s): 2 x 1.25 ml
Application: PCR

Description

Description: qPCR MultiplexMaster highROX is designed for quantitative real-time analysis of DNA samples using Dual Labeled Fluorescent Probes, e.g. TaqMan®, Molecular Beacons or FRET probes. The master mix is specially optimized for setting-up multiplex assays with ≥4 target sequences in a single tube. The system overcomes multiplex limitations of conventional qPCR probe mixes combining an above-average robustness for a multitude of known PCR inhibitors with an excellent sensitivity for amplification of lowest template amounts. The 2x concentrated master mix contains all reagents required for qPCR (except template and primer/probe sets) including a highly processive antibody-inhibited hot-start polymerase and ultra-pure dNTPs. The mixes provide an extremely stringent automatic hot-start allowing reaction set-up and temporary storage at room temperature prior to PCR. The reaction chemistry of the kit is optimized for block-based PCR instruments that are compatible with the evaluation of the ROX reference signal. ROX reference dye: The qPCR MultiplexMaster 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 Dual-labeled DNA probes: 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.

Additional Text

Concentration

2x conc.

Additional Information

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.component||20 µlassay||50 µlassay||final conc. qPCR MultiplexMaster highROX||10 µl||25 µl||1x each primerforward(10 µM)1)||0.6 µl||1.5 µl||300 nM each primerreverse(10 µM)1)||0.6 µl||1.5 µl||300 nM each dual-labeled probe(10 µM)2)||0.4 µl||1 µl||200 nM template DNA||x µl||x µl||

Price
281.00


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