Quantitation of gene expression in neural precursors by reverse-transcription polymerase chain reaction using self-quenched, fluorogenic primers.
AuthorsLowe B, Avila HA, Bloom FR, Gleeson M, Kusser W,
JournalAnal Biochem
PubMed ID12672417
Quantitative RT-PCR using LUX primers was performed to determine the expression patterns of various transcripts in samples of pluripotent, mouse P-19 stem cells. The P-19 cells were used because they transform into neuron-like cells upon retinoic acid treatment. The expression of neural and stem cell genes, including GLUR1, GABA-B1a, NMDA1, ... More
Development of a novel real-time RT-PCR assay with LUX primer for the detection
of swine transmissible gastroenteritis virus.
Real-time RT-PCR assay, based on light upon extension (LUX) fluorogenic
primer and LightCycle technology, was developed for rapid detection of transmissible gastroenteritis
virus (TGEV). Viral RNA from different TGEV isolates and clinical specimens was detected. To evaluate
the sensitivity of the assay, a gel-based RT-PCR method targeted at the same 101 bp sequence ... More
Helicase-dependent isothermal DNA amplification.
AuthorsVincent M, Xu Y, Kong H,
JournalEMBO Rep
PubMed ID15247927
'Polymerase chain reaction is the most widely used method for in vitro DNA amplification. However, it requires thermocycling to separate two DNA strands. In vivo, DNA is replicated by DNA polymerases with various accessory proteins, including a DNA helicase that acts to separate duplex DNA. We have devised a new ... More
Multiplex quantitative PCR using self-quenched primers labeled with a single fluorophore.
'Multiplex quantitative PCR based on novel design of fluorescent primers is described. Fluorogenic primers are labeled with a single fluorophore on a base close to the 3'' end with no quencher required. A tail of 5-7 nt is added to the 5'' end of the primer to form a blunt-end ... More
Effect of primary and secondary structure of oligodeoxyribonucleotides on the fluorescent properties of conjugated dyes.
We studied fluorescence intensity, polarization and lifetime of some commonly used fluorophores conjugated to oligodeoxyribonucleotides with different primary and secondary structures. We found that fluorescence intensity can increase or decrease upon hybridization of the labeled strand to its complement depending on the sequence and position of the fluorophore. Up to ... More