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Agilent Technologies miRNA QPCR Master Mix Instruction Manual

Summary

This essential master mix is designed for precise quantitative PCR (QPCR) detection and analysis of microRNAs (miRNA). The accompanying detailed manual serves as a comprehensive guide, covering reagent preparation, optimal storage conditions, step-by-step amplification protocols, and generating dissociation curves. Ideal for molecular biology researchers in academic R&D settings, it ensures accurate and repeatable miRNA quantification using automated thermal cyclers.

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mi RNA QPCR Master Mix

Instruction Manual Catalog #600583 Revision C

Research Use Only. Not for Use in Diagnostic Procedures. 600583-12

Page Summary Contents For Agilent Technologies miRNA QPCR Master Mix Instruction Manual

Page 1 mi RNA QPCR Master Mix Instruction Manual Catalog #600583 Revision C Research Use Only. Not for Use in Diagnostic Procedures. 600583-12
Page 2 LIMITED PRODUCT WARRANTY This warranty limits our liability to replacement of this product. No other warranties of any kind, express or implied, including without limitation, implied warranties of mer...
Page 3 mi RNA QPCR Master Mix CONTENTS Materials Provided .............................................................................................................................. 1 Storage Conditions ....
Page 4 mi RNA QPCR Master Mix MATERIALS PROVIDED Catalog #600583 Materials Provided Concentration Quantity a mi RNA QPCR Master Mix b 2× 2.5 ml Reference Dye b 1 m M 100 l a The mi RNA QPCR master mix provi...
Page 5 NOTICES TO PURCHASER This product is licensed for research and development only under patents and patent applications jointly owned by BIOTIUM and ALLELOGIC. Use of this product is covered by one or m...
Page 6 INTRODUCTION The mi RNA QPCR Master Mix provides the reagents for quantitative PCR amplification of c DNA templates derived from micro RNAs (mi RNAs) within a total RNA population. Because of their sh...
Page 7 PREPROTOCOL CONSIDERATIONS QPCR Forward Primer Selection The primers used in the QPCR protocol are the universal reverse primer and a unique forward primer that allows specific amplification of the ta...
Page 8 Reference Dye Dilution Recommendations Prepare fresh* dilutions of the reference dye prior to setting up the reactions, and keep all tubes containing the reference dye protected from light as much as ...
Page 9 PROTOCOL QPCR Amplification Notes The master mix is light-sensitive; solutions containing the master mix should be protected from light whenever possible. The protocol for generating c DNA from mi RNA...
Page 10 4. Using c DNA generated from the mi RNA 1st-strand c DNA synthesis kit, add 0.5–1.0 l of c DNA sample to each reaction and gently mix without creating bubbles (do not vortex). Note The optimal quant...
Page 11 TROUBLESHOOTING Observation Suggestion No or low yield of amplification As a positive control, carry out the entire protocol using the Agilent He La-S3 Cell Line product in QPCR Total RNA as the input...
Page 12 REFERENCES 1. Mackay, J. and Landt, O. (2007) Methods Mol Biol 353:237-61. 2. Sang, F. and Ren, J. (2006) J Sep Sci 29(9):1275-80. 3. Kwok, S. and Higuchi, R. (1989) Nature 339(6221):237-8. ENDNOTES A...
Page 13 mi RNA QPCR Master Mix Catalog #600583 QUICK-REFERENCE PROTOCOL 1. If the passive reference dye will be included in the reaction (optional), dilute 1:500 (Mx3005P, Mx3000P, or Mx4000 instruments) or 1...

Manual Details

Brand Agilent Technologies
Pages 13
File Size 194.79 KB
Published May 13, 2026
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Frequently Asked Questions

How many QPCR reactions can I perform with one master mix?

The miRNA QPCR master mix provides enough reagents for two hundred 25-µl QPCR reactions.

What storage temperature should I use after thawing the materials?

After thawing, the master mix may be stored at 4°C for up to six months.

Are there specific primers needed besides the master mix components?

Yes, you need a miRNA-specific forward primer and are recommended to select a QPCR forward primer.

How should I handle the reference dye when preparing reactions?

The master mix and reference dye are light sensitive and must be kept away from light whenever possible.