Yoon OK, Hsu TY, et al. (2012) Genetics and regulatory impact of alternative polyadenylation in human B-lymphoblastoid cells. PLoS Genet, 8(8):e1002882; doi: 10.1371/journal.pgen.1002882.
Yoon et al. examined the regulatory impact of mRNA 3’ untranslated region motifs. The authors performed qPCR experiments using gBlocks® Gene Fragments to generate DNA standard curves for absolute quantification of mRNA in cells transfected with 3’ UTR reporters. This work demonstrates that double-stranded gBlocks Gene Fragments can serve as an ideal source for qPCR standards. They can be ordered up to 3000 bp in length, are normalized to 200–1000 ng, depending on size, and are sequence-verified. Several amplicons can be designed into a single gBlocks Gene Fragment sequence for multiplex amplifications. As an added benefit, because they are limited in quantity and do not require additional processing and purification, gBlocks Gene Fragments can reduce the risk of template contamination in the lab that is common with plasmid DNA.
PrimeTime® qPCR Assays
5′ nuclease, probe-based assays—the gold standard for quantitative gene expression studies
Primer-based assays—designed for intercalating dye experiments
Create custom assays that are designed using our proprietary bioinformatics algorithms for any target and to your specific parameters. Alternatively, select one of our predesigned assays for human, mouse, and rat mRNA targets that are supported by our bioinformatics algorithms and up-to-date sequence/SNP information.
Learn more at www.idtdna.com/PrimeTime. For assistance with assay design, contact our scientific application specialists at email@example.com
ZEN™ and TAO™ Double-Quenched Probes have a 5′ fluorophore, an internal quencher (ZEN or TAO quencher), and Iowa Black® FQ as the 3′ quencher. These probes provide consistently earlier Cq values and improved precision, when compared to traditional, single-quenched qPCR probes.
Learn more at www.idtdna.com/qPCRprobes.
gBlocks® Gene Fragments
gBlocks Gene Fragments are double-stranded, 125–3000 bp DNA molecules. They are ideal for use as qPCR controls and standards, as well as for gene construction and editing applications. These affordable gene fragments are sequence-verified, ship in a few working days, and save laboratory time.
Learn more at www.idtdna.com/gBlocks.
Author: Hans Packer, PhD, is a scientific writer at IDT.
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