PMC:99051 / 13424-14871 JSONTXT

{"target":"https://pubannotation.org/docs/sourcedb/PMC/sourceid/99051","sourcedb":"PMC","sourceid":"99051","source_url":"https://www.ncbi.nlm.nih.gov/pmc/99051","text":"There are 2 variations of the experimental procedure for a given cDNA library (see Fig. 1 and the detailed description in Material and Methods). In (i) a single clone analysis (SCA) the procedure was applied to only one clone at a time, and yielded accurate fingerprints for certain genes experimentally. This did not speed up the process of expression analysis compared to sequencing of clones from cDNA libraries (EST approach), but generated the fingerprints used for the mixed analysis. In (ii) the mixed analyses (MA) the procedure was applied to pools of 96 cDNA clones multiplexing the plasmid preparation, labelling and analysis, thereby speeded up the process by nearly two orders of magnitude. In spite of the multiplexing, already established fingerprints of genes could be unambiguously identified in the restriction fragment mixture derived from the pools (Fig. 1 and 4). If the characteristic fingerprint of a certain gene was identified in the fragment mixture, at least one of the 96 colonies of the pool included a vector with the corresponding cDNA. Therefore, the frequency of occurrence of a fingerprint enabled us to count the abundance of genes and to calculate expression levels from cDNA libraries derived from different statuses at a high throughput. The high throughput only applied to the analysis of the abundance of fingerprints, as in contrast to the SCA, no new fingerprints could be generated by the mixed analysis.","tracks":[]}