gene_expression

PrimeTime® qPCR Primer Assays

Primer-only premixed assays 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 (Molecular Probes) or EvaGreen® (Biotium) dyes. Predesigned assays 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.

  • Advanced bioinformatics delivers the best design possible, with considerations for exon location and number of transcripts detected
  • Achieve >90% efficiency over 4 or more orders of magnitude with our predesigned assays, guaranteed
  • Test target sequence amplification prior to using more expensive probe-based assays
  • Receive primer sequences with all orders

Ordering

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 Primers500$45.00 USD2-3 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

Predesigned qPCR assays 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 $40.00 USD 3 business days 5

PrimeTime qPCR Primer Assays

PrimeTime qPCR Primer Assays are ideal for use with SYBR Green, EvaGreen, 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, and the assays ship in 2–3 business days.

  • 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
  • Ideal for use with SYBR Green, EvaGreen, and other intercalating dyes, where no probe is needed
  • Available in tubes or 96-well plates

Predesigned assays are designed using a proprietary algorithm: in addition to optimized oligo Tm (base composition, oligo length, etc.), the bioinformatic calculations account for factors, such as SNPs (based on current NCBI RefSeq releases), cross-react searches to avoid off-target amplification, recognition of splice variants, and secondary structure predictions.

Guarantee: Predesigned assays are guaranteed to perform with PCR efficiencies of 90–110% and R2 >0.99, or the assay will be replaced with another at no additional charge. Supporting data must be submitted to applicationsupport@idtdna.com.

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 optimized preset design parameters for PCR and qPCR, or customize parameters for your application. The PrimerQuest tool is based on the Primer 3 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 applicationsupport@idtdna.com.

Primer sequences are provided

We provide primer sequences with each order to assist with best practices in research reporting and reproducibility. Specifically, sequence transparency

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

PrimeTime Assays deliver same high efficiency with or without probe

Figure 1. PrimeTime Assays yield the same high efficiency whether used with intercalating dyes or probe. Amplification of 5 sequential 4-fold dilutions of cDNA using PrimeTime qPCR Primer Assays (with SYBR Green dye) 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 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) 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%.

Application guides

Brochures and flyers

Formulation

We can provide custom formulation for PrimeTime qPCR Primer Assays. Contact custcare@idtdna.com for assistance.

GMP and OEM

If you require oligos that are approved for use in molecular diagnostic applications, or if you are interested in our OEM services, please click here.