MGB Eclipse® Probes

MGB Probes -Shorter probes with increased stability and specificity

Traditional Dual Labelled qPCR probes with sequences less than 20 bases will have greater specificity to the target sequence but may struggle to achieve the desired melting temperature (Tm) required within the qPCR reaction.

 

Minor Groove Binding (MGB) Probes utilise a 3’ Eclipse® Dark Quencher coupled to MGB moiety which will bind to Minor Groove of the target sequence and effectively increase the stability of the probe-target duplex. 

 

® Eclipse is a registered trademark of EliTech Group

Increase stability, specificity and SNP Discrimination

Increased stability results in the ability to design shorter, more specific probe designs with higher melting temperature (Tm).

Wide Selection of Flurophores

Including FAM, HEX, ATTO dyes, TexasRed, Yakima Yellow,
JOE, ROX, Cy5

Reduced background fluorescence

The shorter probe length may reduce background fluoresence due closer proximity of the Dye and Quencher.

 

Product Overview

 
01

Scale, yield & method

qPCR probe scale and length

Biolegio three different synthesis scales for qPCR probes oligos: 40 nmol, 200 nmol and 1000 nmol. Prices include synthesis of maximum 40 nucleotides.

 

Yield MGB qPCR probes

 

  • 40 nmol scale:    approx. 5 nmol min. yield
  • 200 nmol scale:   approx. 20 nmol min. yield
  • 1000 nmol scale: approx. 60 nmol min. yield

 

 

 
02

Premixed assays

Order your primes and probes delivered premixed according to your specifications. All fluorophore -quencher combinations are possible, ideal for Real-Time PCR applications.

 
03

Modifications

You can fit your MGB qPCR probe with numerous fluorophores and quenchers from an extensive list of modifications. 

Quencher and dye compatibility

Please follow the link below for a compatibility figure. For multiplexing applications use fluorophores with the least amount of spectral overlap.

>Fluorophores vs Quenchers compatibility
   

How can I fit modifications to my custom MGB qPCR probe?

Select the modifications of your choice in our webshop or in our excel ordering sheet. Select 'MGB probe' in the orderform as the chemistry used for ordering MGB probes.
  

How can I add mixed bases to my oligo?

Use the standard code letters for mixed bases / wobbles. 

 
04

Purifications

All Biolegio qPCR probes are delivered HPLC purified as standard, this is included in the price. 

 
05

Format & concentration

Your qPCR probes can be shipped dry, dissolved or mixed. Standard probes are delivered dissolved in a tube in a concentration of 100 pmol/μl. Custom concentrations are available on request. 

Available buffers

 

  • H20 buffer
  • TE buffer
  • Tris buffer

 
Your primers and probes delivered as a ready-to-use assay?

Probes can be delivered in any format, concentration or combination. Have a look at our oligo dispensing service for more information about this topic.

 
06

Oligo prepaid service

With our oligo prepaid service we make oligo ordering easy. Credit can be easily transferred to your account. Oligos can then be ordered immediately, eliminating possible delays due to purchase authorization. Please contact Biolegio for more information.

 
07

Documentation

All your DNA oligos are delivered with a specification sheet including GC-content, molecular weight, melting temperature (Tm), OD value, sequence and quantity in μgram and nmol. 

 
08

Quality

A stringent quality control system ensures that you can expect the quality of our oligonucleotides to be of the highest standard. Before the synthesis, all chemicals are checked to meet our quality standards. During the fully automated oligo synthesis, all steps of the synthesis are monitored by multiple control functions on our DNA synthesizers. This way we can assure the coupling efficiency of the synthesis to meet our demands.

Synthesis is followed by further quality controls to guarantee the quality of the oligos. All oligos are routinely analyzed by optical density (OD) measurement. In addition, oligos are randomly analyzed by gel electrophoresis and Liquid Chromatography Mass Spectrometry (LCMS). If an oligo does not meet our requirement it’s resynthesized. The result of our stringent QC process is a product of the highest standard. All oligos are delivered with our product specification sheet, which includes: % GC content, yield in nmol and μgram, OD, melting temperature and molecular weight (MW).

  

ISO quality management

 

 

ISO is an independent, non-governmental membership organisation and the worlds largest developer of voluntary international standards. Biolegio’s manufacturing facility is ISO 9001:2015 and ISO 13485:2016 certified. Achieving these certificates ensures good, reliable and safe products as a result of strong product quality and manufacturing procedures and an ongoing development of our quality management system. This ongoing development empowers us to continuously strengthen our ability to identify and exceed the needs and expectations of our customers.