In a recent webinar, Dr. James Willey describes a two color fluorometric assay for more robust quantitation of nucleic acids in highly degraded FFPE and FNA samples. Wiley’s method, described in a recently published article in the journal PLoS One, employs a patented mixture of internal and external standards paired with a pre-amplification step. This system enables measurement of transcription levels of three lung cancer-associated genes and a control gene in clinical samples. All of the fluorometric probes used in the method are labeled with Black Hole Quencher® dyes in BHQplus® probes. The compact design of BHQplus probes is optimal for this application as it confers enhanced sequence discrimination for each target. The webinar takes the audience through assay development, reagent preparation, and the analytical validation experiments.Read More
The BiosearchTech Blog
Single Nucleotide Polymorphisms (SNPs) have been widely studied for over 15 years, and have applications in forensics, disease susceptibility studies, pharmacogenomic and other personalized medicine applications, crop and livestock breeding programs. Numerous genetic methods and companion platforms have been developed for SNP detection encompassing DNA sequencing, microarrays, mass spec, and PCR based techniques. For high sample throughput applications, PCR analysis has emerged as the method of choice for rapid targeted SNP analysis, facilitated by new platforms and reagent cost improvements. On October 30, 2014, Biosearch Technologies sponsored a webinar showcasing the utility of the BHQplus® probe technology for qPCR based SNP analysis. Presenter Cassie Keppel, Laboratory Operations Manager at Douglas Scientific, shared some compelling data showing the accurate and economical utility behind the use of Biosearch’s BHQplus probes on Douglas Scientific’s Array Tape® platform. In this article, we highlight some of Cassie’s findings on the dramatic cost-savings that the BHQplus probe and Array Tape platform combination offers.Read More
Biosearch Technologies has launched qPCRdesign.com, a new microsite targeted specifically for your quantitative PCR needs. In the qPCR Design lab, users can design, analyze, experiment, and play with a variety of interactive tools at their disposal. This is an excellent resource for both new and seasoned qPCR users to access information, advice, or the tools to quickly perform calculations and begin performing assays in no time!Read More
Different oligonucleotide manufacturers deliver probes and primers in a variety of different measured units including moles (e.g. nmol), molarity (e.g. µM), optical density at 260 nm (e.g. OD), and even the number of reactions a scientist can perform. Although these measured units are all technically correct, it is important to understand what each unit is telling you so you know exactly what you are receiving. In this article, we will quickly cover the basics of each oligo delivery unit to help you make the most informed decision when placing your next order for probes and primers.
Quantity in Moles - abbreviated mol, commonly nanomole; (nmol)
This is the default delivery unit Biosearch uses for delivering oligos. The quantity of moles refers to the number of molecules present in the delivered amount. 5 nanomoles (nmol) of one probe will have the same number of molecules as 5 nanomoles (nmol) of another oligo even if the sequence and modifications are totally different. Because the quantity of moles is independent of the physical characteristics and sequence of each oligo compound it makes it a very convenient and transparent unit of measure. Similarly, the use of moles makes it very easy to dilute your oligos to a fixed molar concentration. When oligos are measured in moles, we know that we can compare one oligo to another very easily. This is not the case with units such as OD as we will see below.Read More
The largest recorded Ebola Virus Disease outbreak in history continues to devastate the West African region. This epidemic has presented a public health emergency to national and international disease control centers with aid workers tirelessly working to assist affected communities. Ebola is a single-stranded RNA virus that has five known strains, four of which are pathogenic to humans. The current outbreak is a result of the spread of the Zaire ebolavirus (EBOV) strain which previously averaged a 78% mortality rate 1. In a recent update from the World Health Organization, upwards of 2,600 deaths have been reported as result of the viral infection over the last several months 2.
Don’t waste time performing an extra step. Cut back on pipetting and order a ValuMix™ Assay to have your probes and primers delivered in one tube. The oligos are formulated according to the quantities you specify and delivered dry for maximal stability. ValuMix Assays expedites your research because they:
Do you remember when the most popular method of DNA amplification involved multiple hybridizations, followed by single or 2-primer extension and ligation steps? DNA amplification was a long and tedious process, and if people followed the saying “if it ain’t broke, don’t fix it” the world would certainly be in a different place today. Without rapid PCR assays, it could take several months for medical test results, innocent people would continue to overcrowd prisons, and less research will be achieved but more interns hired (someone needs to perform all those hybridizations).
Topics: Real-time PCR
A "Referral from the Doctor" Blog Article-
Quantitative reverse transcription polymerase chain reaction (RT-qPCR) is a sensitive and
robust method for analyzing RNA. It has been used to corroborate the findings of other genetic methods, such as microarray and RNAse protection assays, and has become the gold standard for quantifying changes in gene expression. Stellaris FISH is a new RNA detection method that provides direct quantification of transcripts in situ, with potential to become the standard for mRNA and lncRNA interrogation using microscopy.
Non-squamous, Non-small cell lung cancer (NSCLC), the most common cause of death in the US and globally, has recently been dealt a significant blow as a result of collaborations among teams from UCSF Thoracic Oncology, Kaiser Permanente Research, and the China Clinical Trials Consortium (CCTC) uniting hospitals and universities across mainland China.
A "Referral from the Doctor" Blog Article-
In qPCR, fluorescent modified oligos may be either linear or non-linear in structure. In either conformation, Förster resonance energy transfer (FRET) combined with the static quenching mechanism inhibits reporter dye emission until hybridization with the target (Read Quenching Mechanisms in Probes to learn more about FRET and static quenching). Upon hybridization the quencher and reporter dye are separated spatially, interrupting the quenching mechanisms and permitting fluorescence emission from the reporter dye. The increase in signal intensity with repeated PCR cycles indicates the accumulation of product and allows for accurate quantification of template. Gene expression analysis by qPCR is dependent upon direct comparisons between the normalized fluorescence emission of individual samples against a standard curve or through ‘relative’ comparison against expression of an internal calibrator gene.
How do I know how bright the dye will be?