Product info

Licence-free MGB Probes for any application

Minor Groove Binder (MGB) Probes are dual-labeled 5' hydrolysis probes consisting of a 5' fluorescent reporter dye and a 3' Eclipse Dark Quencher (EDQ) conjugated to a MGB moiety. Biosearch Technologies offers high quality yet cost effective MGB oligos with a competitive turnaround time and no minimum order quantity.

 

illustration of MGB probe

 

 

Short probe design delivers high specificity and signal-to-noise

The 3' MGB moiety binds non-covalently to the minor groove and stabilises the target-probe duplex, effectively increasing the Tm of the duplex. The improved stability increases the specificity to the target and allows for a shorter probe design.

A shorter probe has greater quenching efficiency because the dye is closer to the quencher. Combined with a non-fluorescent quencher, such as EDQ, the short MGB Probe leads to low background and high signal-to-noise.

Key benefits

  • Short probes with high specificity: The MGB moiety at the 3' end increases the melting temperature (Tm) and stabilises binding, allowing a shorter probe with improved sequence specificity.
  • High signal-to-noise for increased sensitivity: The increased quenching efficiency of MGB Probes improves sensitivity with lower background noise and a higher signal-to-noise ratio.
  • Improved SNP detection: High duplex stability leads to enhanced ∆Tm and improved SNP/mismatch discrimination.
  • Trusted oligo supplier: Biosearch Technologies is one of the most tenured oligo houses. We are known for quality, consistent delivery and flexibility in format.
  • ISO 9001 and ISO 13485 manufacturing options: Experience freedom from the constraints and restrictions of your current oligo supplier by partnering with us from assay design to scale up and commercialisation.

 

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Improved SNP/mismatch discrimination

Single base mismatches have a greater destabilising effect on MGB Probes than other probe types, particularly when the mismatches are within the minor groove binding region. This greatly enhances the ∆Tm for a mismatch and improves SNP discrimination.

 

Trusted supplier of high quality oligos

Our multi-site oligo manufacturing operations provide manufacturing redundancy, risk mitigation and surge capacity. With our unique vertical integration, our chemical manufacturing labs produce most of the critical raw materials used in our own oligo synthesis methods, ensuring a steady and secure supply chain as well as consistent quality at every stage of development. By manufacturing our own dyes, quenchers, specialty amidites, CPG and specialty synthesis columns, we ensure that we are a reliable partner from project inception through commercial scale up.

 

Manufacturing options under ISO 9001 and ISO 13485

We meet your regulatory needs from research and development to scale up and commercialisation with ISO 9001 (RUO grade) and ISO 13485/cGMP (diagnostic grade) certified manufacturing options, as applicable.

 

Dye-quencher options

5' fluorescent dye Abs (nm) Em (nm) 3' quencher
FAM 495 520 EDQ*
TET 521 536 EDQ
CAL Fluor Gold 540 522 544 EDQ
CIV-550 530 550 EDQ
HEX 535 556 EDQ
CAL Fluor Orange 560 538 559 EDQ

*Eclipse Dark Quencher (EDQ)

 

Equivalent performance to industry standard

MGB Probes manufactured at Biosearch Technologies demonstrate equivalent performance to industry-standard MGB Probes.

 

MGB assays figure Figure 1. MGB assays were designed for the E. coli gene lacZ. The probes for each assay were synthesised 5’ FAM, TET, CIV, VIC, HEX or NED and 3’ MGB – Eclipse Dark Quencher (EDQ), by both Biosearch Technologies (blue) and Competitor X (purple). All non-labelled oligonucleotides were synthesised by Biosearch Technologies. The performance of each assay was monitored by running a dilution series (1 x 102 to 1 x 108 copies per reaction) of E. coli genomic DNA in TaqMan™ Fast Advanced Master Mix, following manufacturers recommendations for assay concentrations and cycling conditions. Assays were run on a BIORAD CFX96 (FAM, TET and CIV/VIC) or an ABI QuantStudio5 (HEX/NED). The Cq values for the Biosearch Technologies and Competitor X manufactured assays were compared for each dilution point and shown to have equivalent performance, with PCR efficiency and R2 values passing PCR criteria (efficiency 90-110% and R2 > 0.98) for all dyes.

 

MGB assays synthesised

Figure 2. MGB assays synthesised with 5’ FAM or CIV and 3’ MGB – Eclipse Dark Quencher (EDQ) were designed for maize SNP identification. The cluster plots were generated using maize assays: (A) and (B): PZA03069_4; (C): PZA01688_3; (D): PZA02890_4. Full details of all published assays tested are available on the maize genotyping panel Biosearch Technologies webpage. Assays were run on two different Biosearch Technologies low-volume, high-throughput automation platforms, allowing for liquid handling, thermal cycling and fluorescence detection/analysis. Assays were run on the IntelliQube in 1.6 µL reaction volumes in Array Tape (A and B) and the SNPline workflow in 1 µL reaction volumes in 1536-well plates (C and D). Both crude-extracted (A, C and D) and sbeadex-purified (B) DNA gave clear clusters, allowing for clear and easy genotype identification. All assays were run with Biosearch Technologies’ BHQ Probe Master Mix.

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MGB 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.

 

Similar products

MGB ValuMix assays for qPCR and gene expression offer the convenience of having a custom, dual-labeled MGB probe and primer pair delivered in exact amounts within a single tube at your selected probe-to-primer ratio (between 1:1 to 1:4.5).

MGB ValuMix assays for SNP genotyping include two custom MGB probes and a primer pair delivered at exact quantities within a single tube. Can be ordered as a 40x assay mix.

Order MGB probes and primers delivered each in separate tubes

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Internal modifications are not allowed in standard primers. Please order this sequence as a custom oligonucleotide custom oligonucleotide or click here for help.
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Technical Specs

MGB Probe specifications
PROBE LENGTH 8-30 bases
DYES AVAILABLE FAM, TET, CAL Fluor Gold 540, CIV, HEX, CAL Fluor Orange 560
QUENCHER AVAILABLE Eclipse Dark Quencher (EDQ)
YIELD 10, 20, 60 nmols delivered
PURIFICATION RP-HPLC
DELIVERY FORMAT Dry or in solution (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)

 

MGB ValuMix Assay for qPCR and gene expression
PROBE LENGTH 8-30 bases
DYES AVAILABLE FAM, TET, CAL Fluor Gold 540, CIV, HEX, CAL Fluor Orange 560
QUENCHER AVAILABLE Eclipse Dark Quencher (EDQ)
YIELD 0.5 nmol probe/0.5-2.5 nmol primer (FAM)
5 nmol probe/5-22.5 nmol primer
20 nmol probe/20-90 nmol primer
PURIFICATION Probe – RP HPLC
Primers – salt-free
DELIVERY FORMAT Dry or in solution (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)

 

MGB ValuMix Assay for SNP genotyping
PROBE LENGTH 8-30 bases
DYES AVAILABLE FAM/TET
FAM/CAL Fluor Gold 540
FAM/CIV
FAM/HEX
FAM/CAL Fluor Orange 560
QUENCHER AVAILABLE Eclipse Dark Quencher (EDQ)
YIELD 2 nmol probe/9 nmol primer
5 nmol probe/22.5 nmol primer
12 nmol probe/54 nmol primer
PURIFICATION Probe – RP HPLC
Primers – RPC
DELIVERY FORMAT Dry or in solution (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)

 

Storage conditions

MGB Probes are shipped at ambient temperature. Upon receipt, store at +2 to +8 °C. Once rehydrated, the oligonucleotides should be aliquoted and stored at -30 °C to -15 °C. Multiple freeze-thaw cycles (>10 cycles) should be avoided. Probes should be protected from light.

Reaction volumes and number of reactions per vial

MGB Probes are available in 10, 20, and 60 nmols delivered. Our Reaction Estimator is a useful tool to calculate the number of reactions that can be performed, depending on desired reaction volume and oligonucleotide concentration. For example, 10 nmol of MGB Probe is sufficient for 2,500 x 20 uL reactions containing 200 nM probe.

Oligonucleotide resuspension

Re-suspend the dried probes to make stock solutions. We recommend creating stocks of 100 µM. TE buffer (10 mM Tris, 0.1 mM EDTA, pH 8) is recommended but other molecular biology-grade, nuclease-free diluents may also be used. If another concentration is desired, please see our Oligonucleotide Resuspension Calculator for resuspension assistance.

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