Abstract
This chapter discusses cDNA cloning that constitutes one of the essential steps to isolate and characterize complex eukaryotic genes, and to express them in a wide variety of host cells. Expression cloning has proven extremely powerful if appropriate functional assays or genetic complementation selection systems are available. Successful construction of full-length cDNA libraries depends heavily on the quality of the mRNA preparation. The use of intact, uncontaminated mRNA is essential for generating full-length clones. The use of clean, intact mRNA, fresh RNase-free reverse transcriptase, and a well-prepared vector primer is essential for the efficient synthesis of long cDNA. Pilot transfections with cyclized plasmid should always be undertaken before attempting to make a large library. Expression cloning of transiently expressed cDNAs can be used successfully provided that a sensitive assay is available and that the clones of interest are present in reasonable abundance.
Sign-in/Register to access full text options 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a callDisclaimer: All third-party content on this website/platform is and will remain the property of their respective owners and is provided on "as is" basis without any warranties, express or implied. Use of third-party content does not indicate any affiliation, sponsorship with or endorsement by them. Any references to third-party content is to identify the corresponding services and shall be considered fair use under The CopyrightLaw.
