qPCRBIO Probe Blue Mix Lo-ROX

Art No | PB20.25

qPCRBIO Probe Blue Mix Lo-ROX

Choose size / variation:

from 53.00 CHF

qPCRBIO Probe Blue Mix is a universal probe kit that includes a non-reactive blue dye for easy sample visualisation and enhanced pipetting accuracy.

Description

Whether your application is for a singleplex or multiplex expression study or a diagnostic assay, qPCRBIO Probe Blue Mix is the robust choice for all your probe-based real-time PCR needs.

The universal probe kit is designed to give superior sensitivity and specificity in all probe-based real-time PCR assays, including TaqMan®, Scorpions®, and molecular beacon probes. qPCRBIO Probe Blue Mix contains a non-reactive dye to improve reaction mix visibility and is particularly useful in reducing pipetting errors when loading small volume reactions or using white plates.

Easy-to-see blue mix for greater pipetting precision
High efficiency in multiplex reactions
Rapid extension rate for early Ct values
Market leading sensitivity – increased limit of detection
Efficient amplification from GC and AT-rich sequences
Antibody-mediated hot start technology
Compatible on all real-time PCR platforms – standard and fast cycling conditions

By combining antibody-mediated hot start technology with the latest advances in buffer chemistry we offer market-leading performance with minimal or no optimisation required. qPCRBIO Probe Mix can be used to reliably detect extremely low copy number targets and quantify any DNA template including genomic, cDNA and viral sequences. The enhanced sensitivity of qPCRBIO Probe Blue Mix makes it the perfect choice for multiplexing.

Applications

Absolute quantification
Relative gene expression analysis
TaqMan®, Scorpions® and molecular beacon probes
Detection of extremely low copy number targets
Diagnostic real-time PCR

Use our qPCR Selection Tool to find out which ROX variant is compatible with your instrument.

Storage

The kit should be stored between -30°C and -15°C. Avoid prolonged exposure to light. If stored correctly the kit will retain full activity for 12 months. The kit can be stored at 4°C for 1 month. The kit can go through 30 freeze/thaw cycles with no loss of activity.

Material Safety Data Sheet (PDF)

Product manual (PDF)

Instrument Compatibility Tool

FAQs

Experiment 1

qPCRBIO Probe Blue Mix
a=Singleplex b=Quadplex

Experiment 2

Invitrogen EXPRESS
a=Singleplex b=Quadplex

Experiment 3

Biorad Ssofast Probe Mix
a=Singleplex b=Quadplex

Experiment 1-qPCRBIO Probe Blue Mix a=Singleplex b=Quadplex

Experiment 2-Invitrogen EXPRESS a=Singleplex b=Quadplex

Experiment 3-Biorad Ssofast Probe Mix a=Singleplex b=Quadplex

Experiment 4

qPCRBIO Probe Blue Mix
a=Singleplex b=Quadplex

Experiment 5

Invitrogen EXPRESS
a=Singleplex b=Quadplex

Experiment 6

Biorad Ssofast Probe Mix
a=Singleplex b=Quadplex

Experiment 4-qPCRBIO Probe Blue Mix a=Singleplex b=Quadplex

Experiment 5-Invitrogen EXPRESS a=Singleplex b=Quadplex

Experiment 6-Biorad Ssofast Probe Mix a=Singleplex b=Quadplex

Experiment 7

qPCRBIO Probe Blue Mix Singleplex sensitivity test ACVR2B

Experiment 8

qPCRBIO Probe Blue Mix Singleplex sensitivity test LIMK1

Experiment 9

qPCRBIO Probe Blue Mix Singleplex sensitivity test ACVR1B

Experiment 7-qPCRBIO Probe Blue Mix Singleplex sensitivity test ACVR2B

Experiment 8-qPCRBIO Probe Blue Mix Singleplex sensitivity test LIMK1

Experiment 9-qPCRBIO Probe Blue Mix Singleplex sensitivity test ACVR1B

Figure 1

Experiments 1, 2 and 3 show TaqMan probe amplification traces of human gene ACVR2B in singleplex and in quadplex (ACVR2B, LIMK1, ACVR1B and CDK7) from a cDNA dilution series. a) traces indicate singleplex reactions, b) traces indicate quadplex reactions. qPCRBIO Probe Blue Mix was tested against the latest competitor mixes from Invitrogen (experiment 2) and Biorad (experiment 3). qPCRBIO Probe Blue Mix shows the least PCR inhibition when in multiplex compared to Invitrogen and Biorad mixes. This is evident in more delayed amplification traces in quadplex (b) compared to singleplex (a). Experiments 4, 5 and 6 show TaqMan probe amplification traces of human gene LIMK1 in singleplex and quadplex (ACVR2B, LIMK1, ACVR1B and CDK7). As with experiments 1, 2 and 3, LIMK1 amplification is less inhibited in multiplex in the PCR Biosystems probe mix than the competitor mixes tested. Cycling conditions were 95°C 2min, 40 cycles of 95°C 10sec, 60°C 15sec on Biorad CFX instrument.

Experiments 7, 8 and 9 show TaqMan probe amplification traces from plasmid dilution series of 1×106 copies to 10 copies of DNA. For each gene qPCRBIO Probe Blue Mix amplified with 100% efficiency and detected 10 copies of DNA.

FAQs ROX

ROX (6-carboxy-X-rhodamine) is used as a passive reference dye in ROX-dependent real-time PCR instruments to normalize for variations of fluorescence levels that can arise mainly due to optical path variations among wells. Normalisation of the fluorescence intensity (Rn) is done in real-time PCR software by dividing the emission intensity of the specific signal by the emission intensity of ROX.

ROX does not take part in the PCR reaction and its fluorescence levels are not proportional to the quantity of DNA in each well, so the addition of this fluorophore to a mix provides a constant fluorescent signal during amplification.

Different types of real-time PCR instruments requiring a passive reference standard have different optimal concentrations of ROX, mainly due to the different optical configurations of each system (i.e. the different type of excitation source and optics used).

The addition of either too little or too much ROX would result in a very noisy signal impacting on the results of the reaction. Therefore, it is extremely important for the user to:

Determine the correct ROX concentration to optimise real-time PCR results, and
Check the ROX settings on the software used to set up the reaction

ROX concentrations in qPCRBIO Probe Blue Mixes:

The qPCRBIO Probe Blue Mixes that contain passive reference dyes come in different formulations, each with a different concentration of the passive reference dye:

qPCRBIO Probe Blue Mix Lo-ROX (PB20.25) contains 112 nM ROX
qPCRBIO Probe Blue Mix Hi-ROX (PB20.26) contains 1.12 µM ROX
PCRBIO Probe Blue Mix Separate-ROX (PB20.27) 2x mix contains no ROX and include a separate tube of 50 µM ROX additive. This enables you to choose what concentration of ROX you’d like to use.

Available sizes:

PB20.25-01	100 x 20μL Reactions
PB20.25-05	500 x 20μL Reactions
PB20.25-20	2000 x 20μL Reactions
PB20.25-50	5000 x 20μL Reactions (1 x 50mL bottle)
PB20.25-51	5000 x 20μL Reactions (50 x 1mL in Pouch)