qPCR MultiplexMaster - Model PCR-321 - Master Mix Kit
From Molecular Biology-Real-Time PCR - qPCR Multiplex Mixes
Master mix for multiplex real-time PCR. qPCR MultiplexMaster 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.
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Products Details
For in vitro use only!
- Shipping:shipped on blue ice
- Storage Conditions:store at -20 °C
avoid freeze/thaw cycles
Storage at 4 °C for up to 3 months possible. - Shelf Life:12 months
- Form:liquid
- Concentration:2x conc.
Description
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 mix is optimized for block-based PCR instruments. The mix can also be used with ROX reference dye (#PCR-351) in PCR instruments that are compatible with the evaluation of the ROX signal
Content:
qPCR MultiplexMaster (red cap)
qPCR Pol, dATP, dCTP, dGTP, dUTP, reaction buffer with KCl, (NH4)2SO4, MgCl2 and stabilizers
PCR-grade water (white cap)
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. For amplification of each target sequence a set of two PCR primers and one fluorescent DNA probe that hybridizes to an internal part of the amplicon are required. The sequence of the dual-labeled DNA probe should avoid secondary structure and primer-dimer formation.
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.
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