Process for amplifying and labeling single stranded cDNA by 5amp;prime; ligated adaptor mediated amplification

摘要

This invention relates to a method wherein special adaptors and primers are used for single stranded cDNA amplification. The method comprises contacting RNA with an anchored cDNA synthesis primer which can anneal to RNA, a suitable enzyme which possesses reverse transcriptase activity, and a special designed adaptor which can efficiently be ligated to the 3′ end of single stranded cDNA by T4 DNA ligase. Processes are provided for using specific primers annealing to the adaptors in the 5′ and 3′-end of the single stranded cDNA for PCR amplifying the cDNA. The invention describes the use of this approach for generation of probes starting from low amounts of RNA to be used e.g. in hybridization towards micro-arrays.