Robust real-time PCR master mix for amplification directly from blood, swabs or tissue
2 x conc. master mix
For general laboratory use.
Shipping: shipped on gel packs
Storage Conditions: store at -20 °C
avoid freeze/thaw cycles,
stable at 4 °C for up to 4 weeks
Shelf Life: 12 months
Direct qPCR ProbesMaster 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:
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 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
The mix can also be used in combination with ROX reference dye (#PCR-351) in PCR instruments that are compatible with the evaluation of the ROX signal.
Direct qPCR ProbesMaster (red cap)
2x conc. mix of antibody-blocked Hot Start polymerase, dNTPs, reaction buffer, additives and stabilizers
Extraction Buffer (yellow cap)
Please handle with care and wear personal protective equipment!
PCR-grade Water (white cap)
Before starting, take reagents out from fridge and allow to thaw completely. Vortex all reagents briefly and spin down the liquids.
1. Sample preparation
1.a Blood Samples / Liquid Samples
1.b Samples from nasal or throat swabs
1.c Samples from Animal or Plant Tissue
|Sample size (diameter)
|1x Extraction Buffer
2. Preparation of the PCR Assay
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 below. A reaction volume of 20-50 μl is recommended for most real-time instruments. 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.
|Direct qPCR ProbesMaster
|Forward Primer 11)
|Reverse Primer 11)
|TaqMan® / Dual Labeled Probe 11)
|Forward Primer 22)
|Reverse Primer 22)
|TaqMan® / Dual Labeled Probe 22)
|ROX Reference Dye
|fill up to
|fill up to
Mix the tubes briefly and spin down to remove bubbles.
3. PCR Cycling
Switch on the real-time PCR cycler and set all cycling parameters as recommended in the table below. Place the vials into the instrument and start the program.
Annealing and elongation
60-65 °C 4)
To obtain optimal specificity and amplification results an individual optimization of the recommended parameters is recommended for each particular sample/primer pair.
4. Data Analysis
BIOZ Product Citations: