Zvinhu zvekupindira muPCR maitiro

Munguva yePrcr yekuita, zvimwe zvinhu zvinotsinhira zvinowanzo kusangana.
Nekuda kwekunzwa kwakanyanya kwePCR, kusvibiswa kunoonekwa seimwe yezvakakosha zvinhu zvine chekuita nePCR Mhedzisiro uye inogona kuburitsa mibairo yenhema.
Zvakaenzana zvakakosha ndizvo zvakasiyana-siyana zvinounza kunhema-zvakashata mhedzisiro. Kana imwe kana inokosha zvikamu zvePCr musanganiswa kana kuti implizitified maitiro pachayo anodzivirirwa kana kukanganiswa, iyo denderedzwa leartay inogona kudziviswa. Izvi zvinogona kutungamira mukudzora kugona uye kunyangwe zvisizvo zvakashata mhedzisiro.
Pamusoro pekunzwa, kurasikirwa kweanotarisirwa nucleic acid yekuvimbika inogona kuitika nekuda kwekutumira uye / kana mamiriro ekuchengetedza asati gadzirira. Kunyanya, tembiricha yakakwirira kana yekuchengetedza isina kukwana inogona kutungamira kukuvadzwa kwemasero uye nuclec acids. Sero uye tishu gadziriso uye parafin embirin zvinonyatsozivikanwa zvinokonzeresa zveDNA Sragmentation uye dambudziko rinonyanya kujeka (ona mifananidzo 1 ne2). Mune zviitiko izvi, kunyange kuzviparadzanisa uye kucheneswa hakubatsire.

Mufananidzo 1 | Mhedzisiro yekusaxtilization paDNA DNA
Agarose gel Electrophoresis yakaratidza kuti mhando yeDNA iri rakapatsanurwa kubva kuParafin zvikamu zveAutopsies zvakasiyana. Dhirowa DNA yekare avhareji zvidimbu zvidimbu zvaive zviripo mune zvinobuda zvichienderana nenzira yekugadzirisa. DNA yakachengetedzwa chete kana yakatemwa muNative chando samples uye mune isina kurongeka isina kwayakarerekera. Iko kushandiswa kweiyo yakasimba acidic bouin inogadzirisa kana isina kudzidziswa, mafomu acid-ine formalin yakakonzerwa nekurasikirwa kwakakosha kweDNA. Chidimbu chasara chakakanganisika zvakanyanya.
Kuruboshwe, kureba kwezvimedu kunoratidzwa muKilobase vaviri (KBP)

Mufananidzo 2 | Kupererwa nekuvimbika kweNucleic acid zvinangwa
. Synthesis of DNA ichatoitika pane chidimbu chidiki. Nekudaro, kana iyo primer yekuwedzera nzvimbo iri kushaikwa paDNA Fragment, chete mutsara wekutanga unoitika. Muchiitiko chakaishamisa, zvidimbu zvinogona kutidzora, asi zvibereko zvichave zvidiki uye zviri pasi pezasi mwero.
. Munguva yegungwa rakareba rekudziya
.
Chimwe chinetso chakajairika chinowanzoitika mumanyorerwo emanyorerwo ndewekuongororwa zvishoma-pane-kusunungurwa-kwakakwana kwekutarisa accleic acids kana phenol-chloroform yekuwedzera. Muzviitiko zvakanyanyisa, izvi zvinogona kubatanidzwa nezvenhema zvenhema. Nguva yakawanda inogona kuponeswa neruzha kana enzymatic gaession yemarara marara, asi nzira iyi inowanzoguma nekudzikira prcr kunzwa nekuda kweisina kukwana nucid kuburitswa.

Inhibition yePolymerase chiitiko panguva yekuwedzera

Muzhinji, inhibition inoshandiswa semudziyo wepfungwa kurondedzera zvinhu zvese zvinotungamira kuSuboptimal PRC Mhedzisiro. Mune yakanyanya biochemical sential pfungwa, inhibition inogumira kune chiitiko cheiyo enzyme, ie, inoderedza kushandurwa kwechigadzirwa neiyo yeDNA polymerese.
Zvikamu mumuenzaniso kana akasiyana-siyana ane reagents anogona kunongedza zvakananga kuti enzera
Nekudaro, zvakawanda zvekudyidzana pakati pezvikamu uye zvinangwa-zvine nuclec acids zvakare zvakasarudzwa se 'pcr inthbitors'. Kamwe kutendeka kwesero kukanganiswa nekuzviparadzaniswa uye iyo nuclec acid inoburitswa, kutaurirana pakati pemienzaniso uye mhinduro kwayo yakakomberedza uye chikamu chakasimba chingangoitika. Semuenzaniso, 'scavenger' inogona kusunga kamwe- kana kaviri-yakasimbiswa DNA yekudyidzana uye yekuchenesa nekudzora huwandu hwezvinangwa zvekupedzisira zvichisvika prc reaction.
Muzhinji, PCR Inhbitors iri mumatanho mazhinji emuviri uye reagents anoshandiswa pakuongororwa kweClinsics

Kunyanya prevalent plcristors kunogona kuwanikwa mumabhasitiri uye eukaryotic masero, asiri-tarch Kucheneswa kwema acidsic acids mukati kana mushure mekurwiswa ndiyo nzira yaunosarudza yekubvisa pcr inthbitors.
Nhasi, zvisikwa zvakasiyana-siyana zvekushandisa zvinokatyamadza zvinogona kutsiva akawanda manyoro ebhuku, asi 100% kupora uye / kana kucheneswa kwezvinangwa hazvina kumbobvira zvawanikwa. Zvinogona kuitika zvinogona kuve zviripo mune yakacheneswa nuceic acids kana inogona kunge yakatotora mhedzisiro. Maitiro akasiyana aripo kuti aderedze kukanganisika kweanotenderera. Iko kusarudzwa kweiyo yakakodzera polymerasese kunogona kuve nehunyanzvi hwakakosha pane chiitiko chechiitiko. Dzimwe nzira dzakaratidzwa kuderedza pcr Inhibition dziri kuwedzera polymerase kushamwaridzana kana kushandisa zvinowedzera zvakadai seBSA.
Inhibition yePCR maitiro inogona kuratidzwa nekushandiswa kwemukati maitiro ekudzora (IPC).
Kuchengeta kunofanirwa kutorwa kuti ubvise vese Reagents uye mamwe mhinduro muKitiki, yakadai saTtabol, Edta, Guscn, SDS, kubva kuNhenol, kubva kuNhenol, kubva kuNhenol, kubva kuNhenol, kubva kuNhenol, kubva kuNhenol, kubva kuNhenol, kubva kuNhenel Acid Zvichienderana nekumanikidzwa kwavo, vanogona kuendesa kana kuti inhibit pcr.