Real-time PCR Applications -- Ask TaqMan® Ep. 17 скачать в хорошем качестве

Real-time PCR Applications -- Ask TaqMan® Ep. 17

Submit your real-time PCR questions at https://www.thermofisher.com/us/en/ho... In this video, Sr. Field Applications Specialist Doug Rains covers several possible applications that are possible on Thermo Fisher Scientific real-time PCR instruments. Learn about both quantitative and non-quantitative applications, including those that interrogate DNA, RNA and even protein. Finally, discover convenient options for acquiring high-quality TaqMan® assays for all of these experiment types. Bon giorno, e benvenuti a Ask TaqMan. I'd like to read a question I received recently from Giampiero at the University of Pisa. He asks, "What are the most common applications for real-time PCR?" Well, most notable on the list is looking at relative expression levels of gene targets. Users compare two or more samples -- say, cells from untreated and treated animals -- in order to determine if and how much the expression of their gene of interest is changing. Using Thermo Fisher real-time systems, in combination with either custom or pre-designed assays, users can look at either messenger RNA or small RNA targets, including specific micro RNAs. So what else can a real-time user do with her instrument? So many applications, so little time. A common one is using a standard curve to calculate absolute or relative copy number of a DNA target. This could be a pathogen in food or animal samples, a microbe in water, or any other detectable nucleic acid sequence. Speaking of food, some labs use qPCR to calculate %GMO in food samples in order to ensure that they meet legal standards. Copy number studies on genomic DNA targets are quite important to any number of labs, who typically need either to segregate animals with different copy numbers of a transgene, or to detect and quantify duplication or deletion events associated with a particular phenotype. Keep in mind, I only mention some of the more common possibilities. Fact is, pretty much any application that requires quantifying nucleic acid targets is possible in real-time. So what about experiments that aren't quantitative? The real-time system has you covered there, as well. Perhaps most notable is the allelic discrimination assay, which detects specific single nucleotide polymorphisms in DNA samples, then segregates samples based on their homozygous or heterozygous genotyopes. For researchers wanting to merely detect rather than quantify pathogens, including those present in samples at extremely low levels, our presence/absence application and associated detection reagents are an excellent option. There's also an option on many instruments to run high-resolution melting experiments, which can be used to check percent-methylation of short DNA target regions or to discover small sequence differences among multiple samples. Besides nucleic acid-based work, real-time is increasingly being used to look at proteins. Two applications worth mentioning are Protein Expression technology, which lets users determine relative expression levels of specific protein targets, as well as our protein thermal shift assay, the perfect application for screening different buffer conditions and ligands for their effect on protein thermal stability. And don't forget: regardless of the application, Thermo Fisher offers a full suite of reagents, assays, and analysis software to help you get the job done. That of course includes millions of pre-developed TaqMan Assays for a number of applications, including gene expression, SNP genotyping, and many more.