Under "A Program for Rat Gene Discovery and Mapping" supported by the National Heart, Lung and Blood Institute and the National Eye Institute, University of Iowa researchers have developed over 80 normalized and subtracted cDNA libraries. ESTs were generated from the 5′ end for a nonredundant set of cDNAs, and every EST (3′ and 5′) generated during this project that was uncontaminated and of sufficient quality was annotated with library information and other identified features, and submitted to the dbEST database at the NCBI.
ESTs were generated from the 5′ end for a nonredundant set of cDNAs, leadiner to over 30,000 unique clones being selected and arrayed into the Rat Non-redundant Collection, sequence verified by the University of Iowa.
EST cDNAs were derived from over 100 different adult and embryonic tissues from the Rattus norvegicus strain Sprague-Dawley
Over 30,000 5'-ESTs were selected and arrayed into the Rat Non-redundant Collection
Libraries were constructed using the pT7T3D-Pac vector with a modified polylinker and provide ampicillin resistance. Host used in construction was E. coli DH10B
Why full-insert sequencing is important
EST (expressed sequence tag) clones are not all fully sequenced, only a portion of the cDNA has been sequenced.
Without full-insert sequencing, it is impossible to know whether a clone contains SNPs, alternative splice variants, insertions, or deletions.
Because these EST clones are not fully sequenced, we cannot guarantee clone content.
Baross A et al systematic recovery and analysis of full-ORF human cDNA clones. Genome research 14, 2083 - 2092.
GE Healthcare Dharmacon (“Dharmacon”) is a distributor of multiple gene expression clone collections (cDNAs and ORFs). These clone collections were generated by groups outside of Dharmacon and thus the quality of the collections is largely dependent upon what Dharmacon received from these groups. Specific clone information and plate coordinates were provided to Dharmacon by the suppliers of these clone collections. Dharmacon has not sequence verified each individual clone from these collections. These collections and individual clones are distributed "as is" with no additional product validation or guarantees. Dharmacon has established quality procedures to ensure that individual clones are picked from the identified well in a plate, grown on the correct antibiotic, and are free of phage contamination. Due to the quality of the information provided to Dharmacon, the clone you receive might not match the expected clone. If this occurs, please contact Technical Support (firstname.lastname@example.org)