PCR Loading and Reference Dyes

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PCR Loading and Reference Dyes

Overview

Precision Blue™ Real-Time PCR Dye Workflow

Precision Blue workflow

Pipetting a reaction mix into a PrimePCR assay plate.


 
Consistent performance with and without dye

Precision Blue™ dye allows easy reaction tracking without sacrificing accuracy and precision.

Consistent performance with and without dye

When tested on a PrimePCR™ array plate that contained 91 unique gene expression assays run in duplicate, reactions with and without Precision Blue dye were virtually identical. The average ΔCq value (ΔCq = 0.27) was less than the commonly observed variation of identical technical replicates. Seven genes are shown here. Cq, quantification cycle; RFU, relative fluorescence units.





Precision Blue™ Real-Time PCR Dye

Precision Blue Real-Time PCR Dye

Precision Blue Real-Time PCR Dye is a concentrated, ready-to-use reagent that increases the visibility of real-time PCR reaction mixtures. This unique, advanced formulation facilitates precise pipetting and accurate reaction tracking when loading tubes or plates for real-time qPCR experiments. It is particularly useful when loading white or 384-well plates, which can sometimes be challenging.

Features and Benefits of Precision Blue Real-Time PCR Dye

  • Increase your qPCR accuracy and reproducibility — more consistent reaction volumes reduce technical variation
  • Confirm complete reaction transfer while pipetting — no more wondering if the full reaction volume was transferred out of the tip into the well
  • Decrease setup time without compromising qPCR data quality — work faster but more accurately using visible confirmation to avoid pipetting errors

Precision Blue Dye can be used with both fluorogenic probes and SYBR® Green chemistries and is optimized for use with all Bio-Rad universal real-time supermixes and one-step kits. For ease of use, Precision Blue dye can be added to the full volume of a 2x supermix or one-step kit reaction mix and then stored as indicated. When used as directed, it will not adversely affect PCR specificity, sensitivity, or reproducibility.

Precision Blue dye does not interfere with passive reference dye fluorescence and can be used on both ROX-dependent and ROX-independent instruments. No additional ROX normalization dye is required for ROX-dependent systems when Precision Blue dye is used as instructed.


ROX Passive Reference Dye

ROX Passive Reference Dye

Many real-time PCR thermal cyclers require an internal ROX reference dye for fluorescent signal normalization and correction of well-to-well optical variations. Bio-Rad's ROX Passive Reference Dye allows PCR reagents that do not contain ROX to be used on all ROX-dependent real-time PCR systems.

As the excitation optics of different instrument platforms vary, the optimal ROX reference dye concentration must be matched to each real-time PCR cycler. This ROX reference dye is formulated as a 50x concentrated stock solution for use on the following instruments:

  • ABI PRISM 7000, 7300, 7700, and 7900 systems

This ROX reference dye should be treated as a 750x concentrated solution for systems that use 580–585 nm excitation for passive reference dye, such as the following instruments:

  • Stratagene Mx4000, Mx3000P, or Mx3005P systems
  • ABI PRISM 7500 sequence detection system

Note: An internal ROX reference dye is not required for use with any Bio-Rad real-time PCR detection system.

Related Categories

Supporting Documentation

Online Resources

Storage at –20°C Guaranteed for 12 months at –20°C in a constant-temperature freezer (multiple freeze/thaws not recommended)
Storage at +4°C ROX: guaranteed for 6 months at 4°C (protected from light)
Precision Blue Dye: guaranteed for 3 months at 4°C (protected from light)
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Precision Blue Real-Time PCR Dye
Precision Blue™ Real-Time PCR Dye, 500 x 20 µl reactions, 55 µl

1725555
55 µl, blue visualization dye for use with real-time PCR systems, 500 x 20 µl reactions

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$31.00
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ROX Passive Reference Dye, 0.5 ml

1725858
0.5 ml, passive reference dye for use on ROX-dependent real-time PCR instruments

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$49.00
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Number Description Options
10041731 Precision Blue™ Real-Time PCR Dye Product Insert, Rev A Click to download
10002317 Instruction Manual, ROX Passive Reference Dye, Rev B Click to download
6252 Reagent Comparison Guide for Real-Time PCR Brochure, Rev C Click to download [ Add to Cart (Free) ]
6090 Amplification Reagents and Plastics Brochure, Rev E Click to download [ Add to Cart (Free) ]