PCR – Multimedia

Six major periodontal pathogens were quantified (with standardization to human gDNA) in periodontal pockets of periodontitis patients and healthy subjects by original qPCR assay. Relative amount of pathogens to total bacterial mass increased for P.gingivalis (100-fold), P.intermedia (10-fold) and T.forsythensis (10-fold).

We performed low-volume on-chip DNA typing of single sperm cells isolated by means of laser microdissection. 16plex PCR was successful in 39 of 40 single cell samples yielding a mean PCR efficiency of 62.8%. In addition, we were able to identify a single-cell sample containing more than one cell enabling us to monitor the quality of the whole procedure, and hence, exclude “contaminated” samples from further analysis.

Escherichia coli (E. coli) has been commonly used for the production of biopharmaceuticals. Among the impurities that must be monitored in biopharmaceuticals is residual host-cell DNA (HCD). This study describes the development and subsequent NDA-level validation of a real time PCR procedure developed in response to a client’s need to improve the sensitivity of detection and quantification of residual E. coli HCD in their drug product.

PCR is an essential tool with utility in a variety of advanced applications. To improve the specificity of PCR, a unique approach to "Hot Start" PCR employing primers containing thermolabile modifications has been developed. These modified primers, named CleanAmp™ Primers, are amenable for use in Hot Start activation schemes as the modification is released after an initial denaturation step.

The detection of SNPs (single nucleotide polymorphisms) is an important tool for researchers in order to find mutations in DNA sequences. In this application note we describe a homogeneous fluorescence assay based on PCR. The Amplifluor® SNPs HT Genotyping System from Millipore was used to screen a number of samples with the help of the FLUOstar OPTIMA multidetection microplate reader from BMG LABTECH.

Existing methods for miRNA quantification rely on sequence-dependent probes or chemically modified primers for optimal specificity and often require RNA isolation that is time-consuming labour-intensive and which increase sample variability We have developed a high performance approach for multiplexed detection of mature miRNAs termed modified stem-loop mediated reverse transcription quantitative PCR (mSMRT-qPCR).

Massively parallel DNA sequencing technologies are of pivotal importance in genome biology and medicine, as they can potentially enable comprehensive and systematic evaluation of genetic variation. Currently, these sequencing technologies are geared toward sequencing whole genomes.

Caffeine (1,3,7-trimethyl xanthine), known to have sensory and stimulatory effects, causes adverse effects on health which has resulted in an increased demand for de-caffeinated tea. The transformants obtained by RNAi are being analysed for partial and complete knockdown of one or both the genes using RT-PCR and reduction in caffeine production estimated by HPLC.

The hypothalamic paraventricular nucleus (PVN), an integrating site in the regulation of neuroendocrine and autonomic nervous systems, is composed of heterogeneous neurons, type I and type II, based on their electrophysiological properties. Single cell real-time RT-PCR analysis was made to investigate the molecular basis underlying the properties. The results suggest that Kv1.3, Kv4.2 and Kv4.3 potassium channels are the potential candidates in determining the electrophysiological properties in