When, where, and under what conditions are genes expressed? Scientists are discovering the surprisingly wide range of products of transcription and translation, and how these different expression products determine the growth and health of an organism.
We have developed three methods for sensitive, accurate quantification of m RNA using real-time PCR: Applied Biosystems™ Taq Man® probe–based analysis, Applied Biosystems™ SYBR™ Green dye–based analysis, and digital PCR.
Specifically, the different fluorescent reporter technologies of real-time q PCR are discussed as well as the selection of endogenous controls.
The conceptual framework for data analysis methods is also presented to demystify these analysis techniques.
In this review, we use RNA-seq as an example to bring this topic into focus and to discuss experimental design and validation issues pertaining to next-generation sequencing in the quantification of transcripts.
The next-generation sequencing is a group of new sequencing technologies that are based on randomly amplifying and shotgun sequencing techniques.
The correlation between the results of microarray experiments derived from non-amplified RNA and amplified samples needs to be evaluated in detail.
Xiangqin Cui obtained her Ph D in Genetics in 2001 and did her postdoctoral training in Statistical Genetics from 2001 to 2004.
She is currently an assistant professor in Department of Biostatistics, Section on Statistical Genetics, at the University of Alabama at Birmingham.
Together with your c DNA, these products contain all of the enzymes, buffers, primers and probe necessary to perform SYBR Green real-time PCR analysis.
DNA microarrays are rapidly becoming a fundamental tool in discovery-based genomic and biomedical research.