1. Innovative Algorithms
a) Statistical Optimization: The sophisticated search algorithm calculates all properties of every possible primer and probe within the allowed length and positional boundaries. Every selected primer and probe must meet all specified criteria. It is rejected if any parameter falls outside of its tolerance limits. Beacon Designer™ makes the task of choosing the best possible oligo set as easy as clicking a button. For expert users a list of alternate oligo sets is available and is sortable by several key parameters.
Note: Competitive products use the consecutive screening technique to choose oligos. Possible oligos are screened for each parameter in an arbitrary order for acceptable parameter values and outliers are eliminated in each screen. The result is highly dependent on the order of screening and does not result in the selecting the best oligo.
b) Avoid Cross Homologies: This function is one of the most unique features of Beacon Designer™ and it ensures that the oligo sets designed result in specific amplification and exact detection, BLAST search is run for the templates against the non-redundant database of the organism and the results are interpreted to identify regions of significant cross homologies automatically.
Note: We are not aware of any product that offers this functionality.
c) Default Parameter Selection: Beacon Designer™ provide default parameter values based on considerable research and private communications with several experts and some of the inventors of the qPCR technique. The default parameters work well for most assays. For more challenging assays, complete manual control is available.
Note: This is a
quote from Alex Ryncarz, Epigenomics Inc.,
a long time customer of Beacon Designer™-"Unless
the sequence is exceptionally GC rich or repetitive,
I found that I could simply use the default
parameters. It is not a surprise that some
sequences, especially those shorter than 500
bases and or having a high GC content, do
not provide primer sets or probes with the
default conditions. The inability of the software’s
default conditions to provide primer or probe
designs is an attribute to the consistently
high quality of primer and probe designs that
are produced using Beacon Designer™. Other
design software may also provide good results,
but I have only seen excellent consistency
with Beacon Designer™, which saves both money
2. Many qPCR Assay Technologies
SYBR® Green, TaqMan®, MethyLight, beacons, FRET: Beacon Designer™ enables you to design almost all the popular qPCR technologies including SYBR® Green, TaqMan®, MethyLight, molecular beacons and FRET assays. You can even first test the primers with a SYBR® Green assay before taking the next step of purchasing the more expensive fluorescent labeled probes for any of the three dual labeled technologies.
software offers only the technology the vendor
recommends and expects the user to buy the
reagents from them. Primer Xpress supports
only TaqMan® assays and Light Cycler software
supports FRET exclusively.
NASBA®, a highly sensitive and specific amplification reaction that offers several advantages over other mRNA amplification methods. NASBA® is an isothermal reaction performed at 41C, which obviates the need for a thermal cycler and may facilitate the production of point-of-test devices. A single-stranded antisense RNA product is produced during NASBA®, which can be directly hybridized by a probe sequence to accelerate post-amplification interrogation of the product. In contrast to PCR, background DNA does not interfere with the NASBA® reaction, as single-stranded RNA sequences are specifically targeted. It holds a great promise in diagnostic assay development.
Note: No other
program, to our knowledge, offers NASBA® qPCR
Locked Nucleic Acid or LNA probes are gaining popularity rapidly for SNP detection qPCR Assay because of improved real-time PCR assay success. Incorporation of the LNA bases significantly increases thermal stability, making shorter probes possible for standard reaction conditions.
Note: No other
program, to our knowledge, offers easy to
use design capability for qPCR assays with
LNA substituted probes.
5-methylcytosine is the most frequent covalent base modification of the DNA of eukaryotic cells. The development of MethyLight assay for DNA methylation analysis has significantly helped understanding biological processes such as transcription regulation, genetic imprinting, and tumorigenesis. MethyLight assay accurately determines the relative prevalence of a particular pattern of DNA methylation. The oligonucleotides designed for MethyLight assays allows high degree of specificity, sensitivity, and flexibility for methylation studies.
Using Beacon Designer™, design Scorpions® to achieve sequence-specific priming and PCR product detection using a single oligonucleotide. Scorpions® represent the most recent development in real-time PCR chemistry. The stem-loop format is a common approach for specific detection of PCR products in which the probe element and primer element are physically coupled during the reaction. It is ideally suited for many applications that require rapid detection such as viral load testing and RNA profiling.
7. Multiplex Reactions
Beacon Designer™ helps you design an optimum set of primers and probes for single tube multiplex Taqman® and molecular beacon assays having up to four sequences.
It first designs the best 50 or so primer and probe assays for each sequence. Then the fluorescent probe in each assay is checked for cross dimerization with the primers and probes of all the assays of the other sequences in the reaction. Potential multiplex sets are created by grouping one successful assay for each sequence in all possible combinations. Each multiplex set is then rated according to the formula described earlier, but with the addition of the various cross dimer energy and Tm mismatch terms. The resultant multiplex set rating is a function of the rating of each of the component assays and the compatibility of each assay with the other assays of the multiplex set.
With 50 or so assays for each of the four sequences, there are more than 6 million multiplex sets possible. Each multiplex set has a total of 108 cross compatibility terms in its rating. Thanks to fast computers, efficient programming, and innovative algorithms, nearly a billion interactions are resolved in a flash.
Note: Multiplexing reactions are crucial in the diagnostic assay development, which is one of the most important applications of qPCR technology. No other program, to our knowledge, has the required capability.
8. Continuous Improvement & Unlimited Free Support
We continuously improve the software based on the latest scientific developments and your feedback. We have a team of five software engineers and one biologist for Beacon Designer™ working full time, dedicated to continuous improvement. Telephone, e-mail and fax support for the life of the product free of charge.
9. Versatile Output
You can control what is output. The results are available in a format that can be used in many ways, e.g. synthesis ordering, loading in your own database, printing, sharing results with colleagues.
10. Thousands of Satisfied Customers
Since the first version, Beacon Designer™ has become a premier qPCR design tool for many researchers around the world. The Inventor of Molecular Beacon technology Dr. Sanjay Tyagi uses Beacon Designer™ in his lab. Click here to see what people are saying about Beacon Designer™.
Beacon Designer™ automates the design of real time primers and probes. It is used by molecular biologists worldwide to design successful real time PCR assays. It saves the time and the money involved in failed experiments. Beacon Designer™ is a flexible solution to your real time primer and probe design needs and pays for itself many times over.
You can BLAST search sequences and search for template structures from within the program. The results of both these searches are used while designing primers and probes. The regions that exhibit significant cross homologies and template structures are avoided automatically during the design.