1: Hloov cov khoom siv sim hauv lub sijhawm

Teeb tsa (NTC) kev tswj tsis zoo thiab rov ua nws ntau zaus.Thaum pom tias muaj PCR cov khoom paug hauv chav kuaj, hloov tag nrho cov khoom siv sim hauv lub sijhawm.Xws li: rov dilute thiab npaj cov primers, re-sterilize lub pipette taub hau, EP raj, ddH2O, thiab lwm yam., hloov nrog ib tug tshiab pipette, thiab ib ntus qiv lwm lub chaw soj nstuam los ua PCR thwmsim.Lub chaw kuaj kab mob yuav tsum tau muab tso rau hauv qhov cua thiab irradiated nrog ultraviolet rays kom txog rau thaum cov khoom PCR raug tshem tawm ua ntej yuav mus rau qhov kev sim.

2: Txuas lub sij hawm raug UV

Nws yog tsim nyog sau cia tias kom tshem tawm cov kab mob DNA, cov hluav taws xob ultraviolet yuav tsum tau ncua ntev li 2 teev dua li niaj zaus.Txawm li cas los xij, qhov cuam tshuam ntawm UV irradiation ntawm kev tshem tawm cov khoom me me (hauv qab 200bp) ntawm DNA paug yog tseem tsis zoo.

Ultraviolet wavelength (nm) feem ntau yog 254/300nm, thiab nws yog txaus rau irradiate rau 30 feeb.Nws yuav tsum raug sau tseg tias thaum xaiv UV kom tshem tawm cov kab mob sib kis ntawm cov khoom seem PCR, qhov ntev ntawm cov khoom PCR thiab kev faib tawm ntawm cov hauv paus hauv cov khoom lag luam yuav tsum raug txiav txim siab.UV irradiation tsuas yog siv tau rau cov tawg ntev tshaj 500 bp, thiab tsis tshua muaj kev cuam tshuam rau cov khoom luv luv.

Thaum lub sij hawm UV irradiation, pyrimidine hauv paus hauv PCR cov khoom yuav tsim dimers.Cov dimers no tuaj yeem txiav qhov txuas ntxiv, tab sis tsis yog txhua tus pyrimidines hauv DNA saw tuaj yeem tsim dimers, thiab UV irradiation tuaj yeem ua rau cov dimers..Qhov degree ntawm kev tsim dimer nyob ntawm UV wavelength, hom pyrimidine dimer thiab cov kab ke ntawm nucleotides nyob ib sab ntawm qhov chaw dimer.Yog li, yog tias PCR amplified fragments me me, nws raug nquahu kom siv UNG anti-PCR cov khoom sib kis.

3: Feem ntau siv DNA cov pa phem scavengers

Aerosols tau yooj yim tsim thaum cov pipettes ntxiv, uas tsis yooj yim rau kev zam, thiab nws yuav daws sai sai.Yog li, nws yog qhov tsis xav tsis thoob qhov kev xaiv zoo uas nquag siv tshwj xeeb DNA cov khoom pov tseg los tiv thaiv kev sib kis ntawm DNA ua paug.

4: Siv UNG los tiv thaiv kev ua qias tuaj

Tom qab PCR cov khoom paug raug tshem tawm, lub chaw kuaj sim tuaj yeem siv UNG los tiv thaiv PCR cov khoom kis kab mob los tiv thaiv PCR cov khoom paug.Hauv cov chaw soj nstuam dav dav, koj tuaj yeem ua qhov kev sim yooj yim sib faib, nruj me ntsis cais thaj chaw ntawm PCR cov khoom lag luam los ntawm lwm qhov chaw, tsim kom muaj qee qhov kev sim thiab cov kev cai, thiab ua cov kev cob qhia cuam tshuam los tiv thaiv qhov tshwm sim ntawm PCR cov khoom sib kis.

Cov lus pom zoo: Tsim ib lub chaw kuaj PCR tsim nyog, tswj xyuas qhov chaw PCR zoo, tsim qauv PCR cov txheej txheem kev khiav hauj lwm, thiab tsim kom muaj kev paub txog kev ua haujlwm nruj ntawm cov neeg sim yog cov yuam sij los tiv thaiv lossis txo cov pa phem ntawm PCR kev sim.