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1、MethodsinMolecularBiologyMethodsinMolecularBiologyTMVOLUME221GenerationofGenerationofcDNALibrariescDNALibrariesMethodsandProtocolsMethodsandProtocolsEditedbyEditedbyShao-YaoYingShao-YaoYingHUMANAPRESSHUMANAPRESSComplementaryDNALibraries11ComplementaryDNALibrariesAnOve
2、rviewShao-YaoYing1.IntroductionComplementaryDNAlibrariesreflectgeneexpressionatcertaintimesforspecificcells,whereasgenomicDNAlibrariesrepresentallgeneticinformationinsomaticcells.Thecomplexityofcellularorganizationreflectsageneticprogramthatencodesacollectionofgenesandth
3、emeanstousethembymanufacturingproteinsforcellularstructures,functionalactivities,andreproductionofcellsthemselves.Theessentialaspectofthisprocessisproteinsynthesisbasedontheinformationstoredinthesequenceofnucleotidesthatmakeupagene(atranscribablesegmentofaDNAmolecule)
4、astheblueprint.Theinformationistranscribedasacomplementarysequenceofthenucleotides(mRNAorthetranscript)thatcarriesthegeneticinformationfromthenucleustotheprotein-synthesizingmachineryinthecytoplasm.Then,mRNAistranslatedintothesequenceofaminoacidsthatmakeupaprotein.The
5、basisofthewidelyusednovelstrategiesforthegenerationofcDNAlibrariesarebasepaircomplementarities,reversetranscription,andpolymerasechainreactions.Thischapterpresentssomegeneralinformationontheprinciplesof,biologybehind,basicprotocolsof,andreagentsusedinthegenerationofcD
6、NAlibraries.Hopefully,thisinformationwillhelpresearchersovercomeproblemsencounteredinactualconstructionofcDNAlibraries.1.1.BasePairComplementaritiesNucleicacidsexhibitbasepaircomplementaritiesthatfaithfullyconvertonestrandofRNA/DNAtoacomplementaryone.Althoughallgeneti
7、cinformationFrom:MethodsinMolecularBiology,vol.221:GenerationofcDNALibraries:MethodsandProtocolsEditedby:S.-Y.Ying©HumanaPressInc.,Totowa,NJ1CH01,1-12,12pgs101/03/03,7:31PM2Yinginthesomaticcellsofaspecificorganismcanbeexpressedasatranscript,manyDNAsequencesarenottransc
8、ribed.ThesesegmentsofDNAarethecodingexonsandthenoncodingintrons.Basically,thegeneticinformationisstoredasastrandofaDNAmolecu
