PCR, kana kuti polymerase chain reaction, inzira inoshandiswa kukudza kutevedzana kweDNA.Yakatanga kuvandudzwa muma1980s naKary Mullis, akapihwa mubairo weNobel muChemistry muna 1993 nekuda kwebasa rake.PCR yakashandura molecular biology, zvichiita kuti vatsvakurudzi vawedzere DNA kubva kumasampuli maduku uye kuidzidza zvakadzama.
PCR inhanho-nhanho nhatu inoitika mumhepo inopisa, muchina unogona kukurumidza kushandura tembiricha yemusanganiswa wekuita.Matanho matatu ari denaturation, annealing, uye kuwedzera.
Mudanho rekutanga, denaturation, DNA ine tambo mbiri inopisa kusvika kune tembiricha yepamusoro (kazhinji inotenderera 95 ° C) kuti iparadze mabhondi ehydrogen anobata tambo mbiri pamwechete.Izvi zvinoguma nemamorekuru maviri eDNA ane tambo imwe chete.
Padanho rechipiri, kudzika, tembiricha inodzikiswa kusvika pa55°C kuitira kuti maprimers aenderane kune anowirirana akatevedzana paDNA ine tambo imwe chete.Maprimers zvimedu zvipfupi zveDNA zvakagadzirirwa kuti zvienderane nekutevedzana kwekufarira paDNA inonangwa.
Mudanho rechitatu, kuwedzera, tembiricha inosimudzwa kusvika pa72°C kubvumira iyo Taq polymerase (rudzi rweDNA polymerase) kugadzira tambo itsva yeDNA kubva kune yekutanga.Iyo Taq polymerase inotorwa kubva kubhakitiriya inogara muzvitubu zvinopisa uye inokwanisa kumira tembiricha yepamusoro inoshandiswa muPCR.
Mushure mekutenderera kumwechete kwePCR, mhedzisiro ndeye makopi maviri echinangwa cheDNA kutevedzana.Nokudzokorora matanho matatu ehuwandu hwehuwandu (kazhinji 30-40), nhamba yemakopi echinangwa cheDNA inotevedzana inogona kuwedzerwa zvakanyanya.Izvi zvinoreva kuti kunyange diki diki rekutanga DNA rinogona kukwidziridzwa kuita mamirioni kana kunyange mabhiriyoni emakopi.
PCR ine akawanda maapplication mukutsvaga uye diagnostics.Inoshandiswa mune zvemajini kudzidza kushanda kwemajini uye shanduko, mune zvemberi kuongorora humbowo hweDNA, mukuongororwa kwechirwere chinotapukira kuona kuvepo kwehutachiona, uye mukuongororwa kwepamuviri kuratidza kusakanganiswa kwemajini mu fetus.
PCR yakagadziridzwawo kuti ishandiswe mune akati wandei akasiyana, senge quantitative PCR (qPCR), iyo inobvumira huwandu hweDNA kuyerwa uye kudzoreredza transcription PCR (RT-PCR), iyo inogona kushandiswa kukwidziridza RNA kutevedzana.
Pasinei nekushandisa kwayo kwakawanda, PCR ine painogumira.Zvinoda ruzivo rwekutevedzana kwechinangwa uye dhizaini yeakakodzera maprimers, uye inogona kukanganisa kukanganisa kana mamiriro ekuita asina kugadziridzwa nemazvo.Nekudaro, nekuchenjerera kwekuyedza dhizaini uye kuuraya, PCR inoramba iri imwe yezvishandiso zvine simba mune molecular biology.
Nguva yekutumira: Feb-22-2023