cdna synthese Archives - Gen9 Genetics
The cloning of cDNA, copies of cellular RNA, is one of the classic technologies in molecular biology. Over the past 30 years, cDNA cloning technologies have been enhanced to allow the cloning of large collections of cDNAs, which are critical to today's understanding of the use of genetic information. With the discovery of noncoding RNAs, additional new approaches have been developed for the cloning of short RNAs. However, realizing that much larger portions of genomes are transcribed than anticipated from genome annotations, cDNA cloning faces new challenges in discovering rare transcripts and making corresponding cDNAs available for use. functional studies. This review provides an overview of the current state of cDNA cloning and the possibilities for the discovery and characterization of new RNA families.
CDNA cloning is one of the fundamental technologies in molecular biology, and most of our knowledge about transcripts and proteins is derived from the ability to prepare cDNA copies from RNA and clone them into cDNA libraries. Since the discovery of reverse transcriptases, different protocols have been developed over time for the construction of cDNA libraries. Improvements in library preparation have been instrumental in gene discovery and the creation of large genomic resources. Recent discoveries of new classes of RNA and transcripts expressed at very low levels call for new cDNA cloning approaches to make such RNAs available for functional analysis. Although it is beyond the scope of this article to review all the technical developments of the last 30 years, the key steps in preparing the cDNA library are addressed to highlight the general principles of cDNA cloning (Fig. 1) and to give an overview of the current state. of cDNA cloning and future directions.