cDNA library

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http://youtube.com/watch?v=2nFfxah8RO8

This lecture explains about the cDNA library construction and screening. http://shomusbiology.weebly.com/ • Download the study materials here- • http://shomusbiology.weebly.com/bio-m... • A cDNA library is a combination of cloned cDNA (complementary DNA) fragments inserted into a collection of host cells, which together constitute some portion of the transcriptome of the organism. cDNA is produced from fully transcribed mRNA found in the nucleus and therefore contains only the expressed genes of an organism. Similarly, tissue specific cDNA libraries can be produced. In eukaryotic cells the mature mRNA is already spliced, hence the cDNA produced lacks introns and can be readily expressed in a bacterial cell. While information in cDNA libraries is a powerful and useful tool since gene products are easily identified, the libraries lack information about enhancers, introns, and other regulatory elements found in a genomic DNA library. • cDNA molecules can be cloned by using restriction site linkers. Linkers are short, double stranded pieces of DNA (oligodeoxyribonucleotide) about 8 to 12 nucleotide pairs long that include a restriction endonuclease cleavage site e.g. BamHI. Both the cDNA and the linker have blunt ends which can be ligated together using a high concentration of T4 DNA ligase. Then sticky ends are produced in the cDNA molecule by cleaving the cDNA ends (which now have linkers with an incorporated site) with the appropriate endonuclease. A cloning vector (plasmid) is then also cleaved with the appropriate endonuclease. Following sticky end ligation of the insert into the vector the resulting recombinant DNA molecule is transferred into E. coli host cell for cloning. Source of the article published in description is Wikipedia. I am sharing their material. Copyright by original content developers of Wikipedia. • Link- http://en.wikipedia.org/wiki/Main_Page

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