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1: Tsiva zvigadzirwa zvekuyedza nenguva

nhau812 (1) 

Gadzirisa (NTC) kutonga kwakashata uye dzokorora kakawanda.Kana zvangoonekwa kuti mune PCR chigadzirwa kusvibiswa murabhoritari, tsiva zvese zvekuyedza nenguva.Zvakadai se: wedzerazve uye gadzira maprimers, simbisazve pipette tip, EP chubhu, ddH2O, nezvimwewo, tsiva nepipette itsva, uye kukwereta kwechinguva mamwe marabhoritari kuita PCR kuyedza.Murabhoritari yakasvibiswa ichapinzwa mhepo uye nekuvhenekerwa nemwaranzi yeultraviolet kudzamara PCR chigadzirwa chabviswa chisati chaenderera mberi nekuyedza.

2: Wedzera nguva yekuratidzira UV

nhau812 (2)

Zvakakosha kuziva kuti kubvisa kusvibiswa kweDNA, nguva dzose ultraviolet mwaranzi inofanira kuwedzerwa nemaawa maviri kupfuura nguva dzose.Kunyange zvakadaro, mhedzisiro ye UV irradiation pakubvisa zvidimbu zvidiki (pazasi 200bp) zvekusvibiswa kweDNA hazvisati zvanaka.

Ultraviolet wavelength (nm) inowanzoita 254/300nm, uye inokwana kuita irradiate kwemaminitsi makumi matatu.Izvo zvinofanirwa kucherechedzwa kuti pakusarudza UV kubvisa kusvibiswa kwezvakasara zvigadzirwa zvePCR, kureba kwePCR chigadzirwa uye kugoverwa kwemabhesi mukutevedzana kwechigadzirwa kunofanirwa kutariswa.UV irradiation inongoshanda kune zvimedu zvirefu zviri pamusoro pe500 bp, uye haina mhedzisiro pazvimedu zvipfupi.

Munguva yeUV irradiation, mabhesi epyrimidine muPCR chigadzirwa anoumba dimers.Aya dimers anogona kumisa kuwedzera, asi haasi ese mapyrimidines ari muDNA cheni anogona kuumba dimers, uye UV irradiation inogona zvakare kutyora dimers..Iyo dhigirii yekuumbwa kwe dimer inotsamira pane UV wavelength, mhando yepyrimidine dimer uye kutevedzana kwema nucleotides padhuze nedimer saiti.Naizvozvo, kana PCR yakakwidziridzwa zvimedu zvidiki, zvinokurudzirwa kushandisa iyo UNG anti-PCR chigadzirwa chekusvibisa system.

3:Inowanzo shandiswa DNA kusvibisa scavenger

nhau812 (3)

Aerosols inogadzirwa nyore nyore apo pipettes yakawedzerwa, iyo yakaoma kudzivisa, uye ichagadzirisa nokukurumidza.Naizvozvo, pasina kupokana isarudzo yakanaka kugara uchishandisa yakakosha DNA kusvibisa scavenger kudzivirira kupararira kweDNA kusvibiswa.

4: Shandisa UNG anti-kusvibisa system

nhau812 (4)

Mushure mekunge kusvibiswa kwechigadzirwa chePCR kwabviswa, marabhoritari yekuyedza inogona kushandisa iyo UNG anti-PCR chigadzirwa chetsvina system kudzivirira PCR kusvibiswa kwechigadzirwa.Mumarabhoritari akajairwa, unogona kuita akareruka ekuyedza partitions, kunyatsoparadzanisa PCR chigadzirwa nharaunda kubva kune dzimwe nzvimbo, kumisa mimwe mitemo nemirau yerabhoritari, uye kudzidzisa kwakakodzera kudzivirira kuitika kwekusvibiswa kwePCR chigadzirwa.

Zvinokurudzira: Kugadzira rabhoritari yePCR inonzwisisika, kuchengetedza nharaunda yakanaka yePCR, kugadzira maitiro ePCR anoshanda, uye kusimudzira ruzivo rwakanyanya rwekushanda kwevanoedza ndidzo kiyi dzekudzivirira kana kuderedza kusvibiswa kwePCR kuyedza.


Nguva yekutumira: Aug-12-2021