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Development of 112 unique expressed sequence tags from chicken liver using an arbitrarily primed reverse transcriptase–polymerase chain reaction and single strand conformation gel purification method
Author(s) -
Carré W.,
Diot C.,
Fillon V.,
Crooijmans R. P. M. A.,
Lagarrigue S.,
Morrisson M.,
Vignal A.,
Groenen M. A. M.,
Douaire M.
Publication year - 2001
Publication title -
animal genetics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.756
H-Index - 81
eISSN - 1365-2052
pISSN - 0268-9146
DOI - 10.1046/j.1365-2052.2001.00792.x
Subject(s) - biology , reverse transcriptase , expressed sequence tag , gene , microbiology and biotechnology , polymerase chain reaction , reverse transcription polymerase chain reaction , genetics , synteny , genome , transcription (linguistics) , complementary dna , gene expression , linguistics , philosophy
In order to provide information on chicken genome expression, expressed sequence tags (ESTs) were developed from chicken liver RNAs using a method based on arbitrarily primed reverse transcription–polymerase chain reaction (RT–PCR) of total RNAs. The method is similar to differential display, using one base anchored oligo‐d(T) reverse‐primers and 20‐mer arbitrary forward‐primers. A purification step by single strand conformation gel electrophoresis was added before sequencing. With a ratio of 112 unique sequences out of 155, we found this method to be highly effective when compared with EST production with randomly selected clones from non‐subtracted, non‐normalized libraries. A large proportion of the ESTs sequenced correspond to genes involved in transcriptional and post‐transcriptional events. Cytogenetic mapping was performed for a subset of ESTs and four regions of conserved synteny between chicken and human were confirmed.