Rolling circle amplification-RACE: a method for simultaneous isolation of 5′ and 3′ cDNA ends from amplified cDNA templates

Rolling circle amplification-RACE: a method for simultaneous isolation of 5' and 3' cDNA ends from amplified cDNA templates.

Random-primed cDNA synthesis facilitates the isolation of multiple 5'-cDNA ends by RACE.

The RACE (rapid amplification of cDNA ends) technique (1, 2) can be used to amplify 5'and 3'-cDNA ends that derive from transcripts of low abundance. To isolate the 5' end of a specific cDNA (5' RACE), a small, anti-sense, transcript-specific oligonucleotide is used to prime first-strand cDNA synthesis. The specific first-strand cDNA is then purified, and polyadenylated using terminal deoxynucl...

Simultaneous amplification of 5' and 3' cDNA ends based on template-switching effect and inverse PCR.

A new approach for simultaneously amplifying the 5' and 3' ends of a desired cDNA is described. The method combines the template-switching effect with inverse PCR, which generates the flanking 5' and 3' regions of a certain cDNA with very low or no background. It requires only minimal amounts of total RNA for the synthesis of first-strand cDNA, while the same cDNA can be used to amplify flankin...

Full-length cDNA cloning and determination of mRNA 5' and 3' ends by amplification of adaptor-ligated cDNA.

An efficient cDNA amplification procedure is described for determining of the 5' and 3' ends of mRNAs and cloning full-length cDNAs. In this approach, a double-stranded (ds) adaptor is ligated to both ends of a library of ds cDNA by T4 DNA ligase. This adaptor-ligated ds cDNA is then used to selectively amplify 5'- or 3'-cDNA fragments by PCR with a combination of gene-specific and adaptor-spec...

Optimized rapid amplification of cDNA ends (RACE) for mapping bacterial mRNA transcripts.

A simple, efficient and sensitive RACE-based procedure was developed for the determination of unknown 5' regions from bacterial cDNA. A number of critical modifications were made to the standard RACE method, including the optimization of the RNA extraction, reverse transcription and PCR conditions. This procedure was used to accurately determine the site of transcript initiation and structure o...