文档介绍：MethodsMethods inin MolecularMolecular BiologyBiologyTM
VOLUME 221
GenerationGeneration ofof
cDNAcDNA LibrariesLibraries
MethodsMethods andand ProtocolsProtocols
EditedEdited byby
Shao-YaoShao-Yao YingYing
HUMANA PRESS
Complementary DNA Libraries 1
1
Complementary DNA Libraries
An Overview
Shao-Yao Ying
1. Introduction
Complementary DNA libraries refl ect gene expression at certain times for
specifi c cells, whereas genomic DNA libraries represent all ic information
in somatic cells. plexity of anization refl ects a ic
program that encodes a collection of genes and the means to use them by
manufacturing proteins for cellular structures, functional activities, and
reproduction of cells themselves. The essential aspect of this process is protein
synthesis based on the information stored in the sequence of nucleotides that
make up a gene (a transcribable segment of a DNA molecule) as the blueprint.
The information is transcribed as plementary sequence of the nucleotides
(mRNA or the transcript) that carries the ic information from the nucleus to
the protein-synthesizing machinery in the cytoplasm. Then, mRNA is translated
into the sequence of amino acids that make up a protein. The basis of the
widely used novel strategies for the generation of cDNA libraries are base pair
complementarities, reverse transcription, and polymerase chain reactions.
This chapter presents some general information on the principles of, biology
behind, basic protocols of, and reagents used in the generation of cDNA
libraries. Hopefully, this information will help researchers e problems
encountered in actual construction of cDNA libraries.
. Base plementarities
Nucleic acids exhibit base plementarities that faithfully convert one
strand of RNA/DNA to plementary one. Although all ic information
From: Methods in Molecular Biology, vol. 221: Generation of cDNA Libraries: Methods and Protocols
Edited by: S.-Y.