TeloPrime enables the amplification of full-length cDNA with extreme stringency based on a new innovative technology. It allows more faithful tagging of full-length RNA transcripts that are both capped and polyadenylated compared to any other protocol available.
The new technology of Cap-Dependent Linker Ligation (CDLL) was developed after several years of thorough research and development. The protocol starts with an oligodT priming of the total RNA and long reverse transcription. In the subsequent ligation reaction a double-stranded adapter with a 5’C overhang allows for an atypical base-pairing with the inverted G of the cap structure. By using a double-strand specific ligase, the ligation will only take place if the cap is present and if the RT has really reached the 5’ end of the mRNA. No ligation will take place if no cap is present such as in degraded RNA (low RIN) or if the RT has terminated prematurely because of secondary structure. After second-strand synthesis the full-length ds cDNA is then globally amplified in a PCR reaction using 5’ and 3’ tag specific PCR primers to provide enough material for downstream applications, such as RACE, cloning, sequencing and library or probe generation.
Learn more about TeloPrime Full-Length cDNA Amplification Kit here.