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KSPQ12012 Sigma

KiCqStart® SYBR® Green Primers

Predesigned primers for gene expression analysis

Synonym: Pre-designed RT-qPCR Primers, SYBR Green RT-qPCR Primers

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Description

Application

Product use: gene expression analysis

Features and Benefits

KiCqStart Primers are sensitive, specific, and will save you time and money by eliminating the tedious steps required to develop an effective SYBR Green I RT-qPCR Assay.
Benefits include:
• Target analysis is done
• Design characteristics are set
• Oligo characteristics are set
• Easy to order

General description

Overview
Compatible with any thermal cycler, KiCqStart Primers are ready-to-order, pre-designed primer pairs for quantifying gene expression by SYBR Green I RT-qPCR (Reverse Transcription Quantitative Real-Time PCR). To comply with MIQE, they have been developed using sophisticated bioinformatics tools and validated in silico to avoid off-target amplification1. KiCqStart Primers are available as up to three ranked sets of forward and reverse primer pairs for all genes from common model organisms.

1SNP loci have been avoided during the design process when possible. Please consult dbSNP to determine if your gene has unavoidable SNPs.

To learn more about KiCqStart Primers visit our Frequently Asked Questions section.

Specifications
Design characteristics
• GC Content: 20 - 80%2
• Melting Temperature: 55.0 ± 5 ºC
• Secondary Structure: None
• Excessive 3′ Clamping: Minimal
• Homology: None

Oligo characteristics
• Molecule: DNA (all sequences verified by mass spec QC); no modifications
• Contents: 1 forward and 1 reverse primer
• Length: 18 - 24 bases2
• Purification: RP cartridge
• Format: 3 OD minimum yield, dry; 1 oligonucleotide per tube

Sequences are provided at the time of shipment.

295% of primer pairs fall within these specifications

Rank
Each gene has up to three alternative primer pairs, which are identified as follows:
• 1: The middle of one primer overlaps an exon splice site
• 2: The 5′ or 3′ end of one primer overlaps an exon splice site
• 3: The amplicon, but neither primer, overlaps an exon splice site
• 4: Primer overlap is not possible for various reasons, e.g. the gene has only one exon (primer pairs with this rank may lead to co-amplification of genomic DNA).

Physical properties

Properties include:
• Available species: human, mouse, rat, zebrafish, cow, horse, Arabidopsis, chicken, dog, rabbit, pig, cat, rice, soybean, rhesus, Drosophila, C. elegans, corn, X. laevis, Guinea Pig, and Chinese Hamster.
• Components: two primers, one oligo per tube
• Shipping condition: dry, ambient temperature
• Storage condition: dry or wet single-use aliquots

Legal Information

KiCqStart is a registered trademark used under license.

SYBR is a registered trademark of Life Technologies

Other Notes

Frequently Asked Questions section is available for this product.

Price and Availability

Ordering predesigned KiCqStart SYBR® Green primers for your gene targets is easy using Sigma's state-of-the-art oligonucleotide configuration tool. Open the tool, enter your gene, select your species, choose your primer sets, and add to cart.



All labs need water
Safety & Documentation

Safety Information

Safety Information for this product is unavailable at this time.

Documents

Certificate of Analysis

Protocols & Articles

Articles

Assay Optimization & Validation - PCR Technologies Guide

Assay optimization and validation are essential, even when using assays that have been predesigned and commercially obtained. Optimization is required to ensure that the assay is as sensitive as is r...
Keywords: AGE, Amplification, Buffers, Detection methods, Digestions, Electrophoresis, Gas chromatography, Gel electrophoresis, Gene expression, Melting, Nucleic acid annealing, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymerization reactions, Polymorphisms, Purification, RNA purification, Separation, Size-exclusion chromatography, Transcription

Data Analysis - PCR Technologies Guide

After a traditional PCR has been completed, the data are analyzed by resolution through an agarose gel or, more recently, through a capillary electrophoresis system. For some applications, a qPCR wil...
Keywords: Amplification, Cancer, Capillary electrophoresis, Cell culture, Degradations, Detection methods, Electrophoresis, Gene expression, Growth factors, Indicators, Microarray Analysis, Polymerase chain reaction, Polymerase chain reaction - quantitative, Purification, RNA purification, Sample preparations, Transcription

KiCqStart™ Primers Frequently Asked Questions

What does validated in silico mean? What makes KiCqStart™ Primers MIQE compliant? What is the typical amplicon size? Do KiCqStart Primers detect all splice variants? How do I find out the targeted sp...
Keywords: Polymerase chain reaction, Polymerase chain reaction - quantitative

Protocols

Hot Start Taq Polymerase Protocol to Reduce Non-Specific Amplification

As PCR reactions sit at room temperature, during assay setup, nonspecific amplification can occur via:
Keywords: AGE, Amplification, Electrophoresis, Gel electrophoresis, Gene expression, Nucleic acid annealing, Polymerase chain reaction, Size-exclusion chromatography

Imprint RIP | uRNA Quantitation | MicroRNA primers | MicroRNA Reverse Transcription

Sigma’s Imprint RNA Immunoprecipitation Kit was used to co-purify human argonaute 2 (Ago2)-associated RNAs from HeLa cells, essentially as instructed in the Technical Bulletin. To evaluate the differ...
Carol Kreader
Principal R&D Scientist
Keywords: Buffers, Cell disruption, Immunoprecipitation, Polymerase chain reaction - quantitative, RNA immunoprecipitation, Western blot

Kicqstart SYBR qPCR | Universal SYBR Green qPCR Protocol

KiCqStart SYBR Green qPCR ReadyMix is a 2X concentrated, ready-to-use reaction cocktail that contains all components, except primers and template, for real-time quantitative PCR (qPCR). This unique c...
Keywords: Amplification, Gene expression, Molecular probes, Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Titrations

Quantitative Probe Based PCR for Gene Expression

In recent years Quantitative PCR has reached a level of sensitivity, accuracy, and ease to support use as a routine assay for measuring gene level expression. The field of cancer research is currentl...

SYBR® Green I Dye Quantitative PCR Protocol

Although quantitative PCR uses the same basic concept as traditional PCR, the reactions differ in that the amplicons are generally smaller and are detected indirectly using an additional dye or label...
Keywords: Gene expression, Nucleic acid annealing, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography

Universal SYBR Green Quantitative PCR Protocol

Technology Overview Assay Considerations Methods of Quantification Equipment & Supplies PCR Mix Selection Guide Protocol Troubleshooting Materials References
Keywords: AGE, Amplification, Buffers, Degradations, Electrophoresis, Enzyme activity, Gas chromatography, Gel electrophoresis, Gene expression, Genetic, Melting, Microarray Analysis, Nucleic acid annealing, Nucleic acid denaturation, Nucleic acid hybridization, Polymerase chain reaction, Polymerase chain reaction - quantitative, Polymorphisms, Purification, Sample preparations, Size-exclusion chromatography, Solvents, Titrations

qPCR Reference Gene Selection Protocol

Analysis of gene expression data requires a stable reference or loading control. This reference is usually one or more reference genes. Suitable reference genes are those which are unaffected by diff...
Keywords: Gene expression, Nucleic acid denaturation, Polymerase chain reaction, Polymerase chain reaction - quantitative, Size-exclusion chromatography, Transcription

Related Content

PCR Selection Guide

Sigma-Aldrich offers a wide variety of PCR reagents to meet any experimental needs. Our range of polymerases is customized to meet your End-Point PCR, qPCR, or RT-PCR needs. Our products vary from ro...
Keywords: Polymerase chain reaction, Polymerase chain reaction - quantitative

Peer-Reviewed Papers
15

References

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