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

Consistent performance with and without dye

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

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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, 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

1725555 /templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5555_1389749902.xml 1725555 /default/main/bio-rad/WORKAREA/default 01/14/14 05:13 PM en_US jcv374ir 2017-11-14 07:07:38 true true internet/product_SKU jcv374ir templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5555_1389749902.xml LSR 2018-02-27 06:49:41 /default/main/bio-rad/STAGING N /default/main/bio-rad en 01/14/24 05:17 PM 1517021091271 AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA 2018-02-27 06:49:41 /default/main/bio-rad/WORKAREA/default/templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5555_1389749902.xml templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5555_1389749902.xml 1725555-precision-blue-real-time-pcr-dye-500-x-20-ul-reactions-55-ul 1725555 Precision Blue real-time PCR dye Precision Blue™ Real-Time PCR Dye, 500 x 20 µl reactions, 55 µl 55 µl, blue visualization dye for use with real-time PCR systems, 500 x 20 µl reactions /webroot/web/images/lsr/products/amplification_pcr/sku_thumb/global/precision-blue-real-time-pcr-dye-172-5555-thumb.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_view/global/precision-blue-real-time-pcr-dye-172-5555-detail.jpg Precision Blue Real-Time PCR Dye preview tommy a Precision Blue™ Real-Time PCR Dye Precision Blue™ Real-Time PCR Dye, 55 µl (500 x 20 µl reactions) 1725555, precision blue, blue, dye, visible, tracking, tracking dye, real-time pcr, real-time real-time pcr, realtime, pipetting 01/14/14 05:13 PM 01/14/24 05:17 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA en LSR /LSR/Amplification---PCR/PCR-Reagents N 0 <p>Use Precision Blue Real-Time PCR Dye to increase the visibility of real-time PCR reaction mixtures and facilitate precise pipetting and accurate reaction tracking when loading tubes or plates for qPCR experiments; work faster but more accurately with visible confirmation.</p> <ul> <li>Particularly useful when loading white or 384-well plates</li> <li>Compatible with Bio-Rad Universal Real-Time PCR Supermixes</li> <li>Use with either SYBR<sup>&reg;</sup> Green or fluorogenic probe chemistries</li> </ul> <h2 class="descHeader">Instrument Compatibility</h2> <p>Precision Blue dye has been validated on the following instruments:</p> <ul> <li>Bio-Rad Real-Time PCR Detection Systems: <ul> <li>CFX96&trade;</li> <li>CFX384&trade;</li> <li><a href="/en-us/product/cfx96-touch-real-time-pcr-detection-system">CFX96 Touch&trade;</a></li> <li><a href="/en-us/product/cfx384-touch-real-time-pcr-detection-system">CFX384 Touch&trade;</a></li> <li><a href="/en-us/product/real-time-pcr-detection-systems/cfx-connect-real-time-pcr-detection-system">CFX Connect&trade; real-time PCR detection systems</a></li> </ul> </li> <li>Applied Biosystems 7900HT and ViiA 7 systems</li> </ul> <p>Compatibility on additional systems has been tested but should be validated by the user.</p> <p>Precision Blue Dye 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.</p> <h2 class="descHeader">Related Products</h2> <p>Bio-Rad offers complete workflow solutions for real-time PCR:</p> <ul> <li><a href="/en-us/category/rna-isolation">RNA Isolation Kits</a></li> <li><a href="/en-us/category/pcr-reagents/reverse-transcription-reagents">Reverse Transcription Kits</a></li> <li><a href="/en-us/category/pcr-reagents/supermixes-for-pcr-real-time-pcr">Real-Time PCR Supermixes</a></li> <div id="wetLab"> <script type="text/javascript"><!-- $('#wetLab').load('/webroot/web/html/lsr/products/amplification_pcr/wet-lab.html'); // --></script> </div> </ul> <h2 class="descHeader">Supporting Documentation</h2> <ul> <li>Precision Blue Real-Time PCR Dye <a href="/webroot/web/pdf/lsr/literature/10041731.pdf" target="_blank">Product Insert</a></li> <li>Amplification Reagents and Plastics <a href="/webroot/web/pdf/lsr/literature/Bulletin_6090.pdf" target="_blank">Brochure</a></li> </ul> <h2 class="descHeader">Online Resources<br /></h2> <p>Follow our <a href="/en-us/applications-technologies/qpcr-assay-design-optimization">step-by-step qPCR assay design and optimization protocols</a> to ensure that your qPCR reactions are reproducible and reliable.</p> <p>Learn more about <a href="/en-us/applications-technologies/real-time-pcr-data-analysis">qPCR data analysis</a> and get help with <a href="/en-us/applications-technologies/real-time-pcr-troubleshooting">troubleshooting real-time PCR reactions</a>.</p> /webroot/web/html/lsr/products/SKUs 172-5555.html /webroot/web/html/lsr/products/SKUs/172-5555.html 1 0 false false false /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725555&CartQuote=QT&quantity= false true false $26.00 false null /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725555&AddToDefaultWishList=true&quantity= true false true true false true true true /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725555&quantity= 1725555
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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

1725858 /templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5858_0921120555094.xml 1725858 <?xml version="1.0" encoding="UTF-8"?> <Deployments> <Node Name="QA"> <Deployment Id="StandardMetadata.jcv374kjjcv374kk" Class="com.interwoven.livesite.lscs.model.LSCSDeploymentImpl" Instance="jcv374kjjcv374kk"> <Name>StandardMetadata</Name> <TargetEnviroment>QA</TargetEnviroment> <LastExecutionTime Class="java.util.Date" Month="1" DayOfMonth="26" DayOfWeek="5" Year="2018" Hour="21" Minute="49" Second="45" ZoneOffsetMillis="0" Time="1517003385147">Fri Jan 26 21:49:45 UTC 2018</LastExecutionTime> </Deployment> <Vpath>/default/main/bio-rad/WORKAREA/default/templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5858_0921120555094.xml</Vpath> <LastExecutionTime Class="java.util.Date" Month="1" DayOfMonth="26" DayOfWeek="5" Year="2018" Hour="21" Minute="49" Second="45" ZoneOffsetMillis="0" Time="1517003385147">Fri Jan 26 21:49:45 UTC 2018</LastExecutionTime> </Node> <Node Name="PROD"> <Deployment Id="StandardMetadata.jcv375eljcv375em" Class="com.interwoven.livesite.lscs.model.LSCSDeploymentImpl" Instance="jcv375eljcv375em"> <Name>StandardMetadata</Name> <TargetEnviroment>PROD</TargetEnviroment> <LastExecutionTime Class="java.util.Date" Month="1" DayOfMonth="27" DayOfWeek="6" Year="2018" Hour="2" Minute="42" Second="11" ZoneOffsetMillis="0" Time="1517020931954">Sat Jan 27 02:42:11 UTC 2018</LastExecutionTime> </Deployment> <Vpath>/default/main/bio-rad/WORKAREA/default/templatedata/internet/product_SKU/data/LSR/Amplification---PCR/PCR-Reagents/172-5858_0921120555094.xml</Vpath> <LastExecutionTime Class="java.util.Date" Month="1" DayOfMonth="27" DayOfWeek="6" Year="2018" Hour="2" Minute="42" Second="11" ZoneOffsetMillis="0" Time="1517020931954">Sat Jan 27 02:42:11 UTC 2018</LastExecutionTime> </Node> </Deployments> 09/21/09 12:05 PM en_US jcv374ih 2017-11-14 07:07:38 true true internet/product_SKU LSR N en 09/21/19 12:05 PM 1517021091313 AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA 2017-11-14 07:07:38 1725858-rox-passive-reference-dye-0-5-ml 1725858 ROX Passive Reference Dye ROX Passive Reference Dye, 0.5 ml 0.5 ml, passive reference dye for use on ROX-dependent real-time PCR instruments 0.5 ml, passive reference dye for use on ROX-dependent real-time PCR instruments 0.5 ml, passive reference dye for use on ROX-dependent real-time PCR instruments /webroot/web/images/lsr/products/amplification_pcr/sku_thumb/global/172-5858_tn.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_view/global/172-5858_view.jpg /webroot/web/images/lsr/products/amplification_pcr/sku_feature/global/172-5858_feature.jpg RepEx ROX Passive Reference Dye 0.5 ml, passive reference dye for use on ROX-dependent real-time PCR instruments 1725858, rox, passive, reference, dye, internal, verification, calibration, rox-dependent, dependent, real-time, real, time, pcr, instruments, fluorescent, signal, normalization, and correction, well-to-well, optical, variations, qpcr, reagent 09/21/09 12:05 PM 09/21/19 12:05 PM AE,AI,AL,AM,AR,AT,AU,AZ,BA,BD,BE,BF,BG,BH,BN,BO,BR,BW,CA,CH,CL,CM,CN,CO,CR,CY,CZ,DE,DK,DO,DZ,EC,EE,EG,EH,ER,ES,ET,FI,FM,FO,FR,GA,GE,GF,GH,GP,GR,GT,GU,HK,HN,HR,HT,HU,ID,IE,IL,IN,IS,IT,JM,JO,JP,KE,KH,KR,KW,KZ,LB,LI,LK,LT,LU,LV,MA,MD,MG,MK,ML,MO,MQ,MS,MT,MU,MX,MY,NG,NI,NL,NO,NP,NZ,OM,PA,PE,PF,PG,PH,PK,PL,PR,PS,PT,PW,PY,QA,RO,RS,RU,SA,SB,SE,SG,SI,SK,SN,ST,SV,TG,TH,TN,TO,TR,TT,TW,TZ,UA,UG,UK,US,UY,UZ,VA,VE,VU,XK,YE,ZA en LSR /LSR/Amplification---PCR/PCR-Reagents N 0 <p>Use ROX Passive Reference Dye on real-time thermal cyclers that require an internal reference dye for fluorescent signal normalization and correction of well-to-well optical variations. ROX passive reference dye allows seamless integration of non-ROX-containing PCR reagents on all ROX-dependent real-time instrument systems.</p> <p>Since the excitation optics of different instrument platforms vary, the optimal reference dye concentration must be matched to the given real-time cycler.</p> <p>Use ROX Passive Reference Dye as a 50x concentrated stock solution on the ABI PRISM 7000, 7300, 7700, or 7900 systems. <br /></p> <p>For instruments that use 580&ndash;585 nm excitation for passive reference, such as the Stratagene Mx4000, Mx3000P, or Mx3005P, or the ABI PRISM 7500 sequence detection system, the ROX passive reference dye should be treated as a 750x concentrated solution.<br /></p> <p>An internal reference dye is not required for use with the iCycler iQ<sup>&reg;</sup>, MyiQ&trade;, DNA Engine<sup>&reg;</sup> Opticon 2, or Chromo4&trade; real-time detection systems.</p> <h2 class="descHeader">Related Categories</h2> <ul> <li><a href="/en-us/category/high-fidelity-standard-pcr-reagents">High-Fidelity and Standard PCR Reagents</a></li> <li><a href="/en-us/category/rna-isolation">RNA Isolation Kits</a></li> <li><a href="/en-us/category/pcr-reagents/reverse-transcription-reagents">Reverse Transcription Kits</a></li> <li><a href="/en-us/category/pcr-reagents/supermixes-for-pcr-real-time-pcr">Real-time PCR Supermixes </a></li> <div id="wetLab"> <script type="text/javascript"><!-- $('#wetLab').load('/webroot/web/html/lsr/products/amplification_pcr/wet-lab.html'); // --></script> </div> </ul> <h2 class="descHeader">Supporting Documentation</h2> <ul> <li>ROX Passive Reference Dye <a href="/webroot/web/pdf/lsr/literature/10002317.pdf" target="_blank">Instruction Manual</a></li> <li>Amplification Reagents and Plastics <a href="/webroot/web/pdf/lsr/literature/Bulletin_6090.pdf" target="_blank">Brochure</a></li> </ul> /webroot/web/html/lsr/products/SKUs/autopopulated/ 172-5858_0921120555094_en.html /webroot/web/html/lsr/products/SKUs/autopopulated//172-5858_0921120555094_en.html false false false /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725858&CartQuote=QT&quantity= false true false $49.00 false null /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725858&AddToDefaultWishList=true&quantity= true false true true false true true true /prd/en/US/adirect/biorad?cmd=ajaxAddToCart&AddToCartProductID=1725858&quantity= 1725858
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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
6090 Amplification Reagents and Plastics Brochure Click to download [ Add to Cart (Free) ]

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