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PrimeTime® qPCR probe-based assays

the gold standard for quantitative gene expression studies

 

  • Select from various combinations of dyes and quenchers
  • Choose from 3 reactions scales - 100, 500 or 2500 reactions
  • Order in tubes or 96-well plates

 

PrimeTime qPCR Primer Assays are provided without probe for intercalating dye experiments.

  • Receive 5 nmoles each of forward and reverse primers mixed, sufficient for 500 reactions (20 µL)
  • Order in tubes or 96-well plates

 

 Additional Benefits and Features

  • Primer and probe sequences provided with each order:
    • To assist with compliance with MIQE Guidelines
    • Allow you to fully analyze and potentially troubleshoot your experiments
  • Primer and probe sequences provided with each order:
    • Receive multi-reaction plates, depending on scale ordered - 100, 500, or 2500 reactions [20 µL] per well
    • Order assays with probes that have different dye–quencher combinations in the same plate
    • Generate your own replicate plates for lower cost per reaction
    • Order a minimum of 24 assays or primer pairs per plate

 

PrimeTime® qPCR 5' Nuclease Assays

PrimeTime qPCR 5' Nuclease Assays are offered in three different sizes to meet any qPCR experimental need. In addition, for the Standard and XL sizes, selection of dye–quencher combination and primer-to-probe ratio can be specified to meet unique experimental demands. Assays consist of a forward primer, a reverse primer, and a qPCR probe all delivered in a single tube. Each oligonucleotide undergoes 100% QC by mass spectrometry and all QC results are provided free of charge to the customer on the IDT website.

 

  • Primers and probe mixed and delivered in a single tube or plate well
  • Online assay design tools provide guaranteed performance
  • Available in 5 dye–quencher combinations and 3 reaction scales
  • Choose ZEN™ Double-Quenched Probes in your Assay for superior performance compared to traditional dual-labeled probes
  • MIQE Compliant: All primer and probe sequences are provided
  • Available in tubes or 96-well plates

 

PrimeTime® qPCR 5' Nuclease Assays in Tubes

  No. of Reactions
(20 µL)
Probe
(nmoles)
Primers
(nmoles)1
PrimeTime®
Mini qPCR Assay
100 0.5 1.0
PrimeTime®
Standard qPCR Assay
500 2.5 2.5-10
PrimeTime®
XL qPCR Assay
2500 12.5 12.5-50

 

PrimeTime® qPCR 5' Nuclease Assays in 96-Well Plates

PrimeTime Assay
Plates2
Reactions
(20 µL)
Probe
(nmoles)
Primers
(nmoles)2
PrimeTime®
Assay Plate Mini
100 0.5 1.0
PrimeTime®
Standard qPCR Assay
500 2.5 2.5-10
PrimeTime®
XL qPCR Assay
2500 12.5 12.5-50

1 The primer-to-probe ratio can be specified by the customer (except for PrimeTime Mini).
2 A minimum order of 24 assays is required per plate.

 

Available Dye and Quencher Combinations
for PrimeTime® qPCR 5' Nuclease Assays

5' Dye 3' Quencher Mini Standard XL
FAM ZEN/Iowa Black FQ* point point point
FAM TAMRA   point point
HEX ZEN/Iowa Black FQ*   point point
TET ZEN/Iowa Black FQ   point point
Cy5 Iowa Black RQ   point point

*ZEN™/Iowa Black FQ is a Double-Quenched Probe which provides superior performance to traditional dual-labeled probes.

 

PrimeTime qPCR Primer Assays

 

PrimeTime® qPCR Primer Assays are predesigned qPCR assays used to detect genes in the human, mouse, and rat transcriptomes. These primer sets 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 and normalized to 5 nmoles per primer, and are shipped dried down. Primer sequences are provided upon order, and the primers 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, 96-well plates, or Matrix plates

 

 

PrimeTime® qPCR Primer Assays in Tubes

  Reactions
(20 µL)
Quantity
(nmoles)
PrimeTime qPCR Primers 500 5

 

PrimeTime® qPCR Primer Assays in Plates

  Reactions
(20 µL)
Quantity
(nmoles)
PrimeTime™ Primer Plate 500 5

 

Custom qPCR Probes

Double-quenched qPCR probes

double quenched

While traditional probes have approximately 20–30 bases between the fluorophore and the quencher, the internal ZEN™ or TAO™ quencher decreases that length to only 9 bases. This shortened distance, particularly when combined with the traditional 3’ end quencher, leads to a much more thorough quenching with much lower background and enables the use of much longer probes for designing in AT-rich target regions. In addition to the significantly decreased background, the Double-Quenched Probes also have consistently reduced Cq values and improved precision when compared to traditional probes. Use of the Double-Quenched Probes can allow users to experience both increased sensitivity and precision in their qPCR experiments.

  • ZEN Double-Quenched Probes
    ZEN Double-Quenched Probes contain a 5’ fluorophore; 3’ IBFQ quencher; and proprietary, internal ZEN quencher. The most commonly supplied 5′ fluorophores includes FAM, Hex™, TET™, MAX™, or JOE.
  • TAO Double-Quenched Probes
    TAO Double-Quenched Probes are available with a 5’ Cy® 5 fluorophore; 3’ IBRQ quencher; and proprietary, internal TAO quencher.

Choose from a wide variety of synthesis scales, dyes, and quenchers, including several license-free combinations. Prices and estimated turnaround times depend on the degree of complexity.

 

PrimeTime® qPCR Probes - Standard Level

The minimum guarantees are listed for probes that are 15–35 bases in length.

5' Reporter
Dye(s)
Quencher Synthesis Scale / Minimum Guarantee
FAM ZEN / Iowa Black
FQ *
100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol

 

5' Reporter
Dye(s)
Quencher Synthesis Scale / Minimum Guarantee
FAM TAMRA 100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol
Black Hole
Quencher 1
100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol
HEX ZEN /
Iowa Black FQ *
100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol
Black Hole
Quencher 1
100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol
TET ZEN /
Iowa Black FQ *
100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol
Black Hole
Quencher 1
100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol
YAK ZEN /
Iowa Black FQ *
100 nmol /
15 nmol
250 nmol /
25 nmol
1 umol /
50 nmol

* ZEN/Iowa Black FQ is a double-quenched probe which provides superior performance compared to traditional single-quenched probes.

 

PrimeTime® qPCR Probes - Complex Level

The minimum guarantees are listed for probes that are 15–35 bases in length.

5' Reporter
Dye(s)
Quencher Synthesis Scale / Minimum Guarantee
FAM TAMRA NHS Ester N/A 250 nmol /
8 nmol
1 umol /
20 nmol
MAX NHS ZEN /
Iowa Black FQ *
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 1
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Cy3 Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 2
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
TEX 615 Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 1
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Cy5 Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 2
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
TAO / Iowa
Black RQ *
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
TYE 563 Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 1
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
TYE 665 Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 2
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
JOE NHS Ester Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 1
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
TAMRA NHS
Ester
Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 2
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
ROX NHS Ester Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 1
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Texas Red-X
NHS Ester
Iowa Black RQ 100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol
Black Hole
Quencher 2
100 nmol /
2 nmol
250 nmol /
8 nmol
1 umol /
20 nmol

* ZEN/Iowa Black FQ and TAO/Iowa Black RQ are double-quenched probes which provide superior performance compared to traditional single-quenched probes.

Small-scale qPCR probes

PrimeTime® Mini qPCR Probes

PrimeTime Mini qPCR probes are probes available on a small scale. The smaller scale and lower price makes them ideal for trying out a new probe, for screening the expression levels of many genes, or for testing the conversion to FAM/IBFQ and FAM/ZEN/IBFQ from other FAM-related quenchers. Mini probes are offered with a 5’ FAM and the option of a 3’ IBFQ quencher alone or in combination with the internal ZEN™ quencher. The estimated turnaround time is 3–5 days.

5' Reporter
Dye(s)
Quencher(s) Delivery Amount
FAM ZEN / Iowa Black FQ * 0.5 nmol
 
PrimeTime® Eco qPCR Probes

PrimeTime Mini qPCR probes are probes available on a small scale. The smaller scale and lower price makes them ideal for trying out a new probe, for screening the expression levels of many genes, or for testing the conversion to FAM/IBFQ and FAM/ZEN/IBFQ from other FAM-related quenchers. Mini probes are offered with a 5’ FAM and the option of a 3’ IBFQ quencher alone or in combination with the internal ZEN™ quencher.

5' Reporter
Dye(s)
Quencher(s) Delivery Amount
FAM ZEN / Iowa Black FQ * 2.5 nmol

* ZEN/Iowa Black FQ is a double-quenched probe which provides superior performance compared to traditional single-quenched probes.

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