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PrimeTime™ qPCR Primer Assays

Premixed primer pairs for analyzing gene expression using intercalating dyes.

PrimeTime qPCR Primer Assays provide a primer pair designed for real-time PCR using intercalating dyes, such as SYBR® Green (Thermo Fisher Scientific) or EvaGreen® (Biotium) dyes. Predesigned sequences for human, mouse, or rat are designed with advanced bioinformatic and thermodynamic sequence analytics and for easy selection. Custom assays may be created for any sequence from any species using the PrimerQuest™ Tool.

Ordering

  • Advanced bioinformatics delivers the design you need with considerations for exon location and number of transcripts identified
  • Predesigned sequences will achieve ≥90% efficiency or we will provide an alternative design free of charge
  • Test target sequence amplification prior to using more expensive probe-based assays
  • Receive primer sequences with all orders

PrimeTime qPCR Primer Assays in tubes

Available in standard size, premixed, and shipped dry.


No. of Reactions (20 µL)PriceEstimated Ship DateQuantity (nmoles)
PrimeTime qPCR Primers50041,00 € EUR2-3 business days5

1 Available for human, mouse, and rat targets (for assay configuration, choose intercalating dyes, primers only)

2 Choose design: qPCR–2 primers–intercalating dyes

PrimeTime qPCR Primer Assays in 96-well plates

Custom assays and predesigned qPCR sequences for human, mouse, and rat targets. Available in standard size, premixed, and shipped dry.


ProductReactions (20 µL)PriceEstimated Ship DateQuantity (nmoles)
PrimeTime™ Primer Plate 500 34,32 € EUR 3 business days 5

Product details

PrimeTime qPCR Primer Assays can be used with SYBR® Green (Thermo Fisher Scientific), EvaGreen® (Biotium), and other intercalating dyes, where no probe is needed. PrimeTime qPCR Primer Assays are available in standard size, premixed, normalized to 5 nmol per primer, and shipped dried down. Primer sequences are provided upon order.

PrimeTime qPCR Primer Assays

  • The same primer pairs found in the PrimeTime qPCR 5' Nuclease Assays
  • Forward and reverse primers, mixed and delivered in a single tube or plate well
  • Compatible with SYBR® Green, EvaGreen®, and other intercalating dyes, where no probe is needed
  • Available in tubes or 96-well plates

Predesigned sequences are designed using a proprietary algorithm. The bioinformatic calculations optimizes oligo melting temperature (Tm), base composition, oligo length, etc., and accounts for factors such as SNPs (based on current NCBI RefSeq releases), off-target amplification, splice variants, and secondary structures.

Guarantee: Predesigned sequences will achieve 90% efficiency or better, or we will provide an alternative design free of charge. For more information, please contact us.

Custom primer assays can also be designed for other species using the PrimerQuest™ Tool. This tool may be used to design oligos for endpoint PCR, qPCR, and Sanger sequencing. Use our preset design parameters for PCR and qPCR or customize parameters for your application. The PrimerQuest Tool is based on the Primer3 engine.

For commonly studied pathways in human, mouse, and rat species, we have suggested gene sets (see Resources section below) that can be used with our PrimeTime plate ordering system.

For help with assay design, contact us.

Primer sequences are provided

We provide primer sequences with each order to assist with best practices in research reporting.

  • Boosts publication credibility—you can publish your research according to MIQE guidelines
  • Helps identify and avoid SNPs
  • Facilitates design of multiplex experiments
  • Assists with data interpretation and troubleshooting
  • Allows you to validate your transcripts

Product data

Figure 1. PrimeTime qPCR Primer Assays yield the same high efficiency whether used with intercalating dyes or probes. Amplification of 5 sequential 4-fold dilutions of cDNA using PrimeTime qPCR Primer Assays (with SYBR® Green dye [Thermo Fisher Scientific]) or the PrimeTime qPCR 5′ Nuclease Assay (with dual-labeled probe) targeting human 3-oxoacid CoA transferase 1 (OXCT1) (NM_000436).

Figure 2. PrimeTime qPCR Primer Assays have an average reaction efficiency >90%. Sixty randomly selected PrimeTime qPCR Primer Assays and 15 PrimeTime qPCR Primer Assays for endogenous control genes used with Brilliant III Ultra-Fast SYBR® Green qPCR Master Mix (Agilent) were analyzed over 5 sequential 4-fold dilutions (50–0.195 ng/reaction) of cDNA prepared from Universal Human Reference RNA (Agilent). Reactions were run on the 7900HT Fast Real-Time PCR System (Thermo Fisher Scientific) using PCR cycling conditions: 3 min 95°C; 45 x (5 sec 95°C, 15 sec 60°C). Average reaction efficiencies for the assays tested exceeded 98%.

Resources

Frequently asked questions

*RUO—For research use only. Not for use in diagnostic procedures. Unless otherwise agreed to in writing, IDT does not intend for these products to be used in clinical applications and does not warrant their fitness or suitability for any clinical diagnostic use. Purchaser is solely responsible for all decisions regarding the use of these products and any associated regulatory or legal obligations.

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