Product info

BHQplus™ Probes are compact, dual-labelled, hydrolysis probes for qPCR. This probe type features a 5' fluorescent dye and a 3' Black Hole Quencher™ (BHQ) with the unique BHQplus duplex stabilising chemistry to enhance specificity and mismatch discrimination.

BHQplus Probes use modified C and T nucleotides to stabilise the probe-target duplex, which enables design of shorter oligonucleotides, typically 15 to 25 bases.

Key benefits:

  • Short probes with high specificity: Duplex stabilisers permit the design of shorter dual-labelled probes with increased stability and specificity during hybridization.
  • Ideal for difficult targets: High duplex stability improves SNP/mismatch discrimination and binding to difficult regions including AT-rich targets.
  • Improved assay sensitivity: Our best-in-class BHQ is positioned closer to the dye, resulting in greater quenching efficiency.
  • Versatile: These probes can be paired with a wide range of dyes to fit your exact instrument and application.
 

Greater duplex stability for enhanced specificity and sensitivity

BHQplus Probes form more stable probe-target duplexes, enabling shorter oligonucleotide designs. Shorter oligos offer high specificity and better detection of difficult targets, such as AT-rich sequences. A shorter BHQplus probe will generate lower background and exhibit a higher signal-to-noise ratio, enhancing the sensitivity of your assay.

 

bhqplus melting temp

 

This melting temperature graph shows an example of a BHQplus Probe denaturing at a temperature >10 °C higher than the denaturation of a regular probe, indicating the increased stability of the BHQplus-target duplex.

 

bhqplus melting temp

 

SNP genotyping cluster plot illustrating HotShot extracted maize DNA run with BHQ Probe Master Mix, with maize-specific BHQplus Probe-based assays. The BHQplus Probes were labelled with either FAM (specific for allele X) or CAL Fluor Orange 560 (specific for allele Y). Each data point represents a 1 µL total reaction volume in a 1536-well plate, with the experimental protocol run on the SNPline instrument platform. This experiment returned a 98.9% call rate, out of 96 total samples tested.  This demonstrates high specificity and sensitivity of BHQplus Probes, generating high quality data from crude-extracted DNA on a complex plant species.

 

Achieve optimal quenching efficiency

When it comes to assay design, we don’t believe in a one-dye-fits-all strategy. Our wide selection of dyes empowers you to design the best PCR or qPCR probe for your application without sacrificing performance. We have paired each dye with the BHQ quencher that offers optimal quenching efficiency.

5' fluorescent dye

Abs (nm)

Em (nm)

3' quencher

  FAM 495 520 BHQ-1
  TET 521 536 BHQ-1
  CAL Fluor Gold 540 522 544 BHQ-1
  CIV-550 530 550 BHQ-1*, BHQ-2
  HEX 535 556 BHQ-1
  CAL Fluor Orange 560 538 559 BHQ-1
  CAL Fluor Red 610 590 610 BHQ-2
  Quasar 670 647 670 BHQ-2

* Recommended quencher

Use the spectral overlay tool to view compatible dyes for your instrument. Learn more

Order our premixed ValuMix assays to streamline your experimental setup while reducing pipetting errors

ValuMix assays for qPCR and gene expression

This format contains one custom BHQplus Probe and two primers combined in a single tube.  Adjust primer quantities based on probe amount and select a probe:primer ratio between 1:1 and 1:4.5. 

Key features

  • Probes purified via RP HPLC
  • Salt-free primers
  • Wide selection of dyes: FAM, TET, CAL Fluor Gold 540, CIV-550, HEX, CAL Fluor Orange 560, CAL Fluor Red 610 and Quasar 670
  • Probe quantities: 0.5 nmol (FAM only), 5 nmol, or 20 nmol

ValuMix assays for SNP genotyping

This ValuMix assay format contains two custom BHQplus Probes and two primers combined in a single tube, with a probe to primer ratio of 1:4.5.

Key features

  • Probes purified via RP HPLC
  • Primers purified via RPC
  • Probe 1: FAM
  • Probe 2: Choose from TET, CAL Fluor Gold 540, CIV-550, HEX or CAL Fluor Orange 560
  • Size options: small (2 nmol/probe, 9 nmol/primer), medium (5 nmol/probe, 22.5 nmol/primer) and large (12 nmol/probe, 54 nmol/primer)
  • Available as a 40X solution at 8 μM/probe and 36 μM/primer

Design short BHQplus Probes with ease

Are you designing a new assay or considering BHQplus probes for a sequence currently designed for a different probe technology? For design assistance to achieve optimal results, please contact our assay design service team to redesign your assay with BHQplus probes.

Redesigning your assay for a BHQplus probe is required if you are using a traditional probe sequence that is longer than 25 bases or that has a predicted melting temperature of 70 °C. Alternatively, consider ordering the sequence as a BHQ Probe (traditional length) or BHQnova Probe (extended length).

For design or technical assistance, please contact our assay design and support team with questions regarding probe design.

Order now

BHQplus Probes usually ship in 6 business days   More details

 

  • Oligos for North America, South America, and APAC will be supplied out of our Petaluma, US manufacturing facility while oligos for EMEA will be supplied out of our Lystrup, Denmark manufacturing facility unless you have an agreement in place.
  • Our advertised turnaround time applies to orders received before 12PM for the local manufacturing site. Turnaround time is defined as the time of checkout until shipment.
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This probe sequence is shorter than our recommended criteria for probe length. You may want to consider ordering this sequence as a dual-labeled BHQ probe or BHQplus® probe or MGB probe dual-labeled BHQ probe or BHQplus® probe or MGB probe instead.
You may want to consider ordering an internally quenched probe for sequences 30 bases or longer.
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The internal modification you used in the sequence is not available with this 5' Modification. You need to remove it from your sequence.
Are you trying to include RNA bases? RNA bases are not allowed in standard BHQ probes and so the R character will be treated as a DNA wobble instead (A/G). Please use the custom oligo form custom oligo form to order these modifications or click here for help.
Are you trying to include RNA bases? RNA bases are not allowed in BHQnova probes and so the R character will be treated as a DNA wobble instead (A/G). Please click here for help.
Are you trying to include RNA bases? RNA bases are not allowed in standard BHQplus probes and so the R character will be treated as a DNA wobble instead (A/G). Please use the custom oligo form custom oligo form to order these modifications or click here for help.
Are you trying to include RNA bases? RNA bases are not allowed in standard MGB probes and so the R character will be treated as a DNA wobble instead (A/G). Please use the custom oligo form custom oligo form to order these modifications or click here for help.
RNA bases are not allowed in standard Molecular Beacons. Please use the custom oligo form custom oligo form to order these modifications or click here for help.
Inosine modifications are not allowed in standard BHQ probes. Please order on the custom oligo form custom oligo form or click here for help.
Inosine modifications are not allowed in standard Nova probes. Please order on the custom oligo form custom oligo form or click here for help.
An invalid character has been entered into this sequence. Please order this sequence as a custom oligonucleotide custom oligonucleotide or click here for help.
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Illegal character(s) were entered. Please review your sequence information.
Internal modifications are not allowed in standard Molecular Beacons. Please order this sequence as a custom oligonucleotide custom oligonucleotide or click here for help.
Internal modifications are not allowed in standard BHQplus probes. Please order this sequence as a custom oligonucleotide custom oligonucleotide or click here for help.
Internal modifications are not allowed in standard MGB probes. For additional assistance, click here for help.
Internal modifications are not allowed in standard Nova probes. Please order this sequence as a custom oligonucleotide custom oligonucleotide or click here for help.
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This oligo contains a number of 'wobbles'. Please consider limiting this to one trinucleotide wobble and one dinucleotide wobble in two different locations or three dinucleotide wobbles in three different locations such that there is a maximum of 6 variants in a single oligonucleotide.
Inosine modifications must be included in brackets such as [I] or [i].
Internal modifications are not allowed in standard primers. Please order this sequence as a custom oligonucleotide custom oligonucleotide or click here for help.
No special modifications are allowed in BHQplus® probes and primers. Use the custom oligo order form custom oligo order form to order sequences with specialty modifications or click here for help.
No special modifications are allowed in MGB probes and primers. Use the custom oligo order form custom oligo order form to order sequences with specialty modifications or click here for help.
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This oligo contains a number of 'wobbles'. Please consider limiting this to one trinucleotide wobble and one dinucleotide wobble in two different locations or three dinucleotide wobbles in three different locations such that there is a maximum of 6 variants in a single oligonucleotide.
Inosine modifications must be included in brackets such as [I] or [i].
Internal modifications are not allowed in standard primers. Please order this sequence as a custom oligonucleotide custom oligonucleotide or click here for help.
No special modifications are allowed in BHQplus® probes and primers. Use the custom oligo order form custom oligo order form to order sequences with specialty modifications or click here for help.
No special modifications are allowed in MGB probes and primers. Use the custom oligo order form custom oligo order form to order sequences with specialty modifications or click here for help.
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Technical Specs

BHQplus Probe specifications
Probe length 15-25 bases
Dyes FAM, TET, CAL Fluor Gold 540, CIV-550, HEX, CAL Fluor Orange 560, CAL Fluor Red 610, Quasar 670
Quenchers BHQ-1, BHQ-2
Yield 10, 20, or 60 nmols delivered
Purification Reverse Phase HPLC (RP HPLC)
Delivery format Dry or in solution (water, TrisHCl, T10E0.1, T10E1)
Quality control MS and UHPLC
Quality standard ISO 9001 or ISO 13485
Shelf life 12 months from date of manufacture
Shipping conditions Ambient temperature or dry ice (in solution)

 

BHQplus ValuMix for qPCR and gene expression
Probe length 15-25 bases
Dyes FAM, TET, CAL Fluor Gold 540, CIV-550, HEX, CAL Fluor Orange 560, CAL Fluor Red 610, Quasar 670
Quenchers BHQ-1, BHQ-2
Yield 0.5 nmol probe / 0.5-2.5 nmol primer (FAM only)
5 nmol probe / 5-22.5 nmol primer
20 nmol probe / 20-90 nmol primer
Purification Probes - Reverse Phase HPLC (RP HPLC)
Primers - salt-free
Delivery format Dry or in solution (water, TrisHCl, T10E0.1, T10E1)
Quality control MS and UHPLC
Quality standard ISO 9001
Shelf life 12 months from date of manufacture
Shipping conditions Ambient temperature or dry ice (in solution)

 

BHQplus ValuMix for SNP genotyping
Probe length 15-25 bases
Dyes FAM / TET
FAM / CAL Fluor Gold 540
FAM / CIV-550
FAM / HEX
FAM / CAL Fluor Orange 560
Quenchers BHQ-1, BHQ-2
Yield 2 nmol probe / 9 nmol primer
5 nmol probe / 22.5 nmol primer
12 nmol probe / 54 nmol primer
Purification Probe - Reverse Phase HPLC (RP HPLC)
Primers - Reverse Phase Cartridge (RPC)
Delivery format Dry or in solution (water, TrisHCl, T10E0.1, T10E1)
Quality control MS and UHPLC
Quality standard ISO 9001
Shelf life 12 months from date of manufacture
Shipping conditions Ambient temperature or dry ice (in solution)

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