An efficient strategy to isolate full-length cDNAs based on an mRNA cap retention procedure (CAPture).

AUTOR(ES)

Edery, I

RESUMO

The ability to generate cDNA libraries is one of the most fundamental procedures in contemporary molecular biology. One of the major drawbacks of current methods is that most cDNAs present in any given library are incomplete, rendering the characterization of genes an inefficient and time-consuming task. We have developed an affinity selection procedure using a fusion protein containing the murine cap-binding protein (eukaryotic initiation factor 4E), coupled to a solid support matrix, that allows for the purification of mRNAs via the 5' cap structure. When combined with a single-strand-specific RNase digestion step, specific retention of complete cDNA-RNA duplexes following first-strand synthesis is achieved. This method can be used to generate cDNA libraries in which polyadenylated and nonpolyadenylated mRNAs are equally represented and to enrich for full-length or 5'-end clones, thus facilitating cDNA cloning and promoter mapping.

Documentos Relacionados

• Antisense transcripts with rice full-length cDNAs
• Protein–Protein Interaction Panel Using Mouse Full-Length cDNAs
• Full-length cDNAs from chicken bursal lymphocytes to facilitate gene function analysis
• DBTSS: DataBase of human Transcriptional Start Sites and full-length cDNAs
• Splice Variation in Mouse Full-Length cDNAs Identified by Mapping to the Mouse Genome