Ukuseka umfuniselo we-PCR we-SOP ukumisela umgangatho wokuziphatha kwabasebenzi bovavanyo.
Umfunisi uthobela ngokungqongqo iinkqubo zokusebenza, kwaye unciphisa ungcoliseko lwePCR olunokuthi lubangelwe yimiba yabantu okanye luthintele ukwenzeka kongcoliseko.Ukongeza, umntu ovavanyayo kufuneka abe nolwazi oluhambelanayo lobuchwephesha kunye nezakhono, kubandakanya nobuchule kwizixhobo ezinxulumene nokusebenza, ukucacisa yonke inkqubo yomsebenzi, ukuqonda iindlela zonyango zongcoliseko kunye neendlela zolawulo lomgangatho welebhu, kunye nokukwazi ukutolika ngokuchanekileyo iziphumo zovavanyo.
Ukuseka ilebhu ye-PCR eqhelekileyo.
Ilabhoratri ye-PCR yahlulwe yaba ziindawo ezine ngokomgaqo, ezizezi, indawo yokulungiselela i-reagent, indawo yokusetyenzwa kwesampulu, indawo yokukhulisa, kunye nendawo yohlalutyo lwemveliso.Iindawo ezimbini zokuqala ziindawo zokwandiswa kwangaphambili, kwaye iindawo ezimbini zokugqibela ziindawo ezisemva kolwandiso.Indawo yokwandiswa kwangaphambili kunye nommandla we-post-amplification kufuneka yohlulwe ngokungqongqo.Izinto zovavanyo, iirejenti, iphepha lokurekhoda, iipeni, izinto zokucoca, njl.njl., zinokuhamba kuphela ukusuka kwindawo yokwandiswa kwangaphambili ukuya kwindawo yasemva kokukhulisa, oko kukuthi, ukusuka kwindawo yokulungiselela i-reagent → indawo yokusetyenzwa kwesampuli → indawo yokukhulisa → indawo yohlalutyo lwemveliso yokukhulisa, kwaye akufuneki ibuye umva .I-airflow kwi-laboratory kufuneka iphinde ihambe ukusuka kwindawo yokwandiswa kwangaphambili ukuya kwindawo ye-post-amplification, kwaye ingabuyeli ngasemva.Uyilo olufanelekileyo lwelabhoratri yePCR luboniswe ngezantsi:
Umzobo A: Imo ye-PCR efanelekileyo yokuseta ilabhoratri enoxinzelelo olubi kwigumbi le-buffer
Umzobo B: Imo ye-PCR efanelekileyo yokuseta ilabhoratri enoxinzelelo oluhle kwigumbi le-buffer
Imizobo yokuseta elabhoratri ye-PCR enikwe kuMfanekiso A kunye noMzobo B kufuneka ibe yeyona ndlela ifanelekileyo yokuseta, kwaye ilabhoratri enemiqathango ingabhekisa kule mowudi yoyilo.Kwiilabhoratri eziqhelekileyo, kucetyiswa ukuba indawo yokukhulisa i-PCR kunye nendawo yokuhlalutya imveliso inokwahlulwa, kwaye ukuvulwa kwekhava kufuneka kuncitshiswe kangangoko kunokwenzeka kwindawo yokulungiselela isampula kunye nendawo yokukhulisa i-PCR.Khumbula: Iimveliso kunye nezibonelelo zovavanyo kwindawo yohlalutyo lwemveliso zithintelwe ngokungqongqo ukuba zisiwe kwindawo yolungiselelo lwesampulu kunye nendawo yokukhulisa i-PCR.
Ukuba ilabhoratri ifumanisa kuphela i-PCR kunye nokuchonga, kuyacetyiswa ukuba kusetyenziswe i-fluorescent quantitative PCR endaweni ye-PCR eqhelekileyo.
Fluorescence quantitative PCR iziphumo zokufumanisa zingaqokelelwa kwaye zihlalutywe ngeempawu ze-fluorescence, ngoko akukho mfuneko yokuvula isiciko se-electrophoresis emva kokusabela, ethintela ukungcoliswa kwemveliso ye-PCR ebangelwa ukuvuza kweemveliso zokusabela ukwenza i-aerosols.Ukuba unyusa inani lokuvuleka kwe-cap ngexesha lokulayisha i-gel electrophoresis, ukungcoliswa kwe-aerosol kunokwenzeka.Kucetyiswa ukukhuthaza ukusetyenziswa kwe-PCR yobungakanani kwaye ngokuthe ngcembe ithathe indawo ye-PCR yomgangatho.
Inkqubo yokungcoliseka kwemveliso ye-UNG isetyenziselwa ukusabela kwe-PCR.
Inkqubo isebenzisa i-dUTP endaweni ye-dTTP.Emva kokusabela kwe-PCR, zonke iimveliso ze-PCR (amaqhekeza e-DNA) zifakwe kunye ne-dUTP;kumjikelo olandelayo wokusabela kwe-PCR, i-enzyme ye-UNG eyongeziweyo kwinkqubo ifakwe kwi-37 ° C imizuzu emi-5 ngaphambi kwe-PCR, enokuthi ithobe ngokukodwa zonke iziqwenga ze-DNA eziqulethe i-dUTP, kwaye emva koko yenze i-PCR reaction.Oku kunokususa ngokupheleleyo ungcoliseko lwe-aerosol olubangelwa ziimveliso zePCR.Isiphumo siboniswe kulo mfanekiso ungezantsi:
Qaphela: Kuluhlu oluthe ngqo lwe-PCR, ungakhetha uthotho lweemveliso zenkqubo yokungcoliseka kwemveliso ye-PCR ye-Foregene.Cebisa
Kwiilabhoratri ezenza uvavanyo olukhulu lwe-genotyping, kucetyiswa ngamandla ukuba kusetyenziswe inkqubo yokungcoliseka kwemveliso ye-UNG ye-PCR yokuvavanya ii-reagents ukongeza kulwakhiwo lweelabhoratri ezifanelekileyo.
Isikhumbuzo: Usetyenziso lwale nkqubo alunakukususa ungcoliseko lwemveliso yePCR esele yenziwe.Ke ngoko, inkqubo ye-UNG kufuneka isetyenziswe ekuqaleni kovavanyo olufanelekileyo, kwaye inkqubo ye-UNG kufuneka isetyenziselwe ukukhulisa i-PCR, ukuze kuthintelwe ukungcoliseka kweemveliso ze-PCR.
Kuyacetyiswa ukuba usebenzise inkqubo ye-PCR-UNG ye-Foregene xa uqhuba uvavanyo olukhulu, olunje:
Isityalo seLeaf Direct PCR Kit-UNG;
Imbewu yokutyala ngokuthe ngqo kwi-PCR Kit-UNG;
I-Animal Tissue Direct PCR Kit-UNG;
Mouse Umsila Direct PCR Kit-UNG;
I-Zebra Fish Direct PCR Kit-UNG.
Olu luhlu lweekiti ezivela kwi-Foregeneayinakukwazi ukwenza ubhaqo lwe-PCR ngokukhawuleza nakwizinga elikhulu, kodwa ikwakhusela ngokusebenzayo kwaye ilawule ukungcoliseka kwemveliso ye-PCR.
Ixesha lokuposa: Mar-19-2021
