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Kuberekwa kwePCR

PCR (Polymerase Chain Reaction)

Pave nemakore anopfuura makumi matatu kubva pakugadzirwa kwepolymerase chain reaction.Kweanopfuura makore makumi matatu, mushure mekunge nyanzvi dzakawanda pasirese dzichiramba dzichiwedzera nekuvandudza, tekinoroji yePCR yave iyo inonyanya kushandiswa uye inowanzoshandiswa uye yakakosha nzira yekutsvagisa muchikamu chese cheHupenyu Sayenzi.

Iyo TouchDown PCR, Real-Time PCR, Multi PCR, nezvimwewo yakagadziridzwa pahwaro hwekushandiswa kwakakura kwechinyakare PCR tekinoroji, pamwe neiyo ichangoburwa Digital PCR (digital PCR), yakapfumisa zvikuru nzira dzekutsvagisa dzeruzhinji rwevaongorori vesainzi uye nekukasira zvakanyanya hurongwa hwekuvandudza hwemazuva ano Hupenyu Sayenzi, kunyanya molecular biology, yaita tsvakiridzo yehupenyu hwese uye hupenyu hwese.

PCR-musimboti
Polymerase-Chain-Reaction-PCR

Kukanganisa kwechinyakare PCR tekinoroji

Complex nucleic acid kupatsanurwa uyekutorwa:

★ Traditional PCR tekinoroji: inodiwa

★ PCR yakabva teknolojia: inodiwa

★ DNA uye RNA samples: misiyano mikuru, zvakaoma kushanda zvinodiwa

★ Njodzi dzemuviri: zvinhu zvine chepfu zvinokuvadza muviri

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Chinyakare PCR tekinoroji uye tekinoroji inotorwa ine prerequisite-nucleic acid kupatsanurwa uye kucheneswa.

Chero sampu yebhayoloji inoda kuenda kuburikidza neyakatevedzana uye inonetesa sampuli yekugadzirisa kuti uwane nucleic acid samples dzinosangana nezvinodiwa zvePCR tekinoroji.

Kupatsanurwa uye kubviswa kweDNA neRNA kwagara riri basa rekutanga iro rinofanirwa kutsvagirwa nesainzi rinofanira kudzokorora zuva rega rega.

Nekuda kwekusiyana kukuru pakati pemasampuli, kuparadzaniswa uye kuburitsa maitiro eDNA neRNA zvakare akasiyana zvakanyanya.Iri basa rinoda hunyanzvi hwepamusoro hwevashandisi.Kuparadzaniswa kwechinyakare uye nzira dzekubvisa dzinoda kubata kwenguva refu nemamwe mareagents emakemikari ane muchetura.Izvo zvinokonzeresa kukuvadzwa kusinga gadzirise kumuviri wemushandisi, uye kunyange kukonzera kukuvadza kwakananga panguva yekuyedza.

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Panguva imwecheteyo, kune avo vane nhamba huru yemasampuli ekudzidza, kupatsanurwa uye kubviswa kwe nucleic acids ibasa rinonyanya kushanda.

Nucleic acid yekuzviparadzanisa uye yekubvisa kits pamusika ikozvino yakura uye kune akawanda mabhureki, asi akafanana akafanana.Kunyangwe iri silica gel membrane column centrifugal kit kana kuti magnetic bead method kit, inotora nguva yakawanda uye inodhura.Pamusoro pemutengo wekiti, kune zvakare zvakakosha zvinodiwa zvemidziyo yerabhoritari.Iyo automated workstation inoshandiswa mune iyo magineti bead nzira ndeye yakajairwa yakakura-yepamusoro-yakakosha midziyo, inova muripo wakakura werabhoritari.

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Muchidimbu

Usati waita bvunzo dzePCR, pretreatment yemasampuli chinhu chisingadzivisiki uye chinogara chiri musoro kune vanoongorora.Maitiro ekugadzirisa dambudziko iri uye kana PCR kuyedza inogona kuitwa pasina kupatsanurwa uye kutorwa kwe nucleic acids kwagara kuri kufunga kweruzhinji rwevaongorori vesainzi uye vashandi vekiriniki laboratory.

Foregene's Solution

Mushure memakore ekutsvagisa kunorwadza nezve Direct PCR tekinoroji nemamwe makiti ane hukama, Forgene akabudirira kupaza mabhodhoro mazhinji uye akabudirira kuwana yakananga PCR yemhando dzakawanda dzemasamples akasiyana ane kuramba kwakasimba uye kuchinjika, zvichibvumira vaongorori kubvisa kuomarara uye kune njodzi kupatsanurwa uye kubviswa kwe nucleic acids.Izvi zvichaderedza zvakanyanya kuwanda kwekushanda kwemunhu wese, kukurumidzira maitiro ekuyedza, uye kuchengetedza tsvakiridzo yesainzi uye mitengo yekuyedza.

Kunzwisisa kwaForgene uye ruzivo rweDirectPCR

Chekutanga, DirectPCR tekinoroji ndeye yakananga PCR tekinoroji kune akasiyana biological sample tissues.Pasi peiyi tekinoroji mamiriro, hapana chikonzero chekuparadzanisa uye kubvisa nucleic acids, uye sampuli yetishu inoshandiswa zvakananga sechinhu, uye iyo inotarirwa gene primers inowedzerwa kuPCR reaction.

Chechipiri, DirectPCR tekinoroji haingori yechinyakare DNA template amplification tekinoroji, asi zvakare inosanganisira RNA template reverse transcription PCR.

Chechitatu, tekinoroji yeDirectPCR haingoite zvakanangana nemaitiro ePCR pamasampula ematishu, asi inosanganisira Real-Nguva qPCR maitiro, ayo anoda kuti maitiro ekuita ive nesimba rakasimba rekuramba kupindira kwefluorescence uye kupikisa endogenous fluorescence quenchers.

Chechina, mashizha emagetsi anotarirwa nekombiyuta yeDirectPCR inongoda kusunungurwa kwe nucleic acid templates uye usabvisa mapuroteni, polysaccharides, salt ions, nezvimwewo zvinokanganisa maitiro ePCR.Izvi zvinoda iyo nucleic acid polymerase uye PCR Musanganisi mune yekupindura sisitimu kuti ive yakanakisa anti-kudzoserwa uye kuchinjika, uye inogona kuve nechokwadi chekuita enzyme uye kudzokorora kwechokwadi pasi pemamiriro akaomarara.

Chechishanu, masampula ematishu anonangwa neiyo DirectPCR tekinoroji haana kuitiswa chero nucleic acid inopfumisa kurapwa, uye huwandu hwetemplate hudiki kwazvo, hunoda kunzwisiswa kwakanyanya uye kukwidziridzwa kwemaitiro ekuita.

Mhedziso

DirectPCR tekinoroji ndeimwe yeakanyanya kukosha kuvandudza tekinoroji uye hutsva mumakore makumi matatu apfuura kubva pakuzvarwa kwePCR tekinoroji.Forgene ane uye acharamba ari piyona uye muvambi weiyi tekinoroji.

Iyo tarisiro yekushandisa yeDirectPCR tekinoroji yakakura kwazvo.Kuenderera mberi kwekuvandudza uye kusimudzirwa kweiyi tekinoroji zvechokwadi kuchaunza shanduko dzekupandukira kutsvagiridzo yesainzi nebasa rekuongorora.Uku ndiko shanduko yePCR tekinoroji.


Nguva yekutumira: Feb-21-2017