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
✍ Scribed by W. Carré; C. Diot; V. Fillon; R. P. M. A. Crooijmans; S. Lagarrigue; M. Morrisson; A. Vignal; M. A. M. Groenen; M. Douaire
- Publisher
- John Wiley and Sons
- Year
- 2001
- Tongue
- English
- Weight
- 127 KB
- Volume
- 32
- Category
- Article
- ISSN
- 0268-9146
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✦ Synopsis
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.