Absolute quantification of the budding yeast transcriptome by means of competitive PCR between genomic and complementary DNAs.

ABSTRACT:

Background

An ideal format to describe transcriptome would be its composition measured on the scale of absolute numbers of individual mRNAs per cell. It would help not only to precisely grasp the structure of the transcriptome but also to accelerate data exchange and integration.

Results

We conceived an idea of competitive PCR between genomic DNA and cDNA. Since the former contains every gene exactly at the same copy number, it can serve as an ideal normalization standard for the latter to obtain stoichiometric composition data of the transcriptome. This data can then be easily converted to absolute quantification data provided with an appropriate calibration. To implement this idea, we improved adaptor-tagged competitive PCR, originally developed for relative quantification

SUBMITTER: Miura F 

PROVIDER: S-EPMC2612024 | biostudies-literature | 2008 Nov

REPOSITORIES: biostudies-literature