I-Polymerase chain reactions (PCR) ingenye yezindlela ezaziwa kabanzi ezisetshenziswa kulabhorethri yesayensi yempilo.
Amapuleti e-PCR akhiqizwa kumapulasitiki ekilasi lokuqala ukuze acutshungulwe kahle kakhulu futhi ahlaziywe amasampula noma imiphumela eqoqiwe.
Zinezindonga ezincane nezifanayo ukuze zinikeze ukudluliswa kwe-thermal okunembile.
Ukulungiselela izinhlelo zokusebenza zesikhathi sangempela, ingxenye yamaminithi ye-DNA noma i-RNA iyavalwa futhi igcinwe kumapuleti e-PCR.
Amapuleti e-PCR asebenza kahle kakhulu ekuvaleni ukushisa futhi anqanda ukugeleza kokushisa futhi.
Kodwa-ke, njengoba amapuleti e-PCR esebenza futhi enokwethenjelwa, amaphutha nokungalungi kuyancipha kalula ngenkathi kusetshenzwa amasampula.
Ngakho-ke, uma unentshisekelo yokuthola ikhwalithi enhle nephezuluAmapuleti e-PCR.Kuhle ukuxhumana nomkhiqizi wamapuleti we-PCR onokwethenjelwa. Ngalokhu uyaqiniseka ngokuthola isivumelwano esihle kakhulu.
Nazi ezinye izindlela zokuphepha okufanele uzilandele ukuze ugweme ukungcoliswa kwama-reagents noma amasampula futhi uvimbele ukungalungi ukuthi kungene emiphumeleni.
Ukuvala inzalo ezindaweni ezikuzungezile
Okuhle noma okungalungile kwenzeka ngenxa yokuba khona kokungcola, okwenza ungabaze imiphumela.
Ukungcola nokungcola kwenzeka ngezindlela ezihlukahlukene njenge-DNA engahlobene noma izithasiselo zamakhemikhali ezigcina zinciphisa ukusebenza kahle nokusebenza kokusabela.
Kunezindlela eziningi zokunciphisa kakhulu izinga lokungcoliswa kwepuleti le-PCR.
Ukusebenzisa amathiphu okuhlunga amagciwane kungenye indlela ewusizo yokuvimbela ukungcola ukuthi kungene kumasampula akho ngamapayipi.
Nikeza isethi yemishini ehlanzekile ngokuphelele, ehlanganisa ama-pipette nama-rack, ukusetshenziswa kwe-PCR kuphela. Lokhu kuzoqinisekisa ukudluliswa okungenandaba kokungcola noma ukungcola endaweni yokusebenzela.
Sebenzisa ama-bleach, i-ethanol kuma-pipette, ama-racks namabhentshi ukuze usule ukungcola.
Nikeza isikhala esibekelwe konke ukusabela kwakho kwe-PCR ukuze uqhubeke unciphisa ukungcoliswa kwezinhlayiyana.
Sebenzisa amagilavu ahlanzekile kuzo zonke izinyathelo futhi uwashintshe njalo.
Amapuleti e-PCR
Hlola Ukugxila Nokuhlanzeka Kwesifanekiso.
Ukuhlanzeka kwebhentshi nezinto zokusebenza ezisetshenziswa lapho kuhlaziywa amasampula nge-PCR kufanele kugcinwe. Kubalulekile ukuqinisekisa izinga lokuhlanzeka kwamasampuli ngaphambi kokuhlaziywa nokucutshungulwa.
Ngokuvamile, abahlaziyi bacabangela izinga lokugxila kanye nokuhlanzeka kwamasampula e-DNA.
Zama isilinganiso sokumunca okungu-260nm/280nm akumele sibe ngaphansi kuka-1.8. Ngenkathi ubude begagasi obulandelayo obuphakathi kuka-230nm no-320nm busetshenziselwa ukukhomba ukungcola.
Kwesinye isikhathi, usawoti we-chaotropic nezinye izinhlanganisela eziphilayo zitholwa ngezinga lokumunca elingu-230nm. Ngenkathi uthuthuva kumasampula e-DNA nawo ayatholakala futhi aqinisekiswa ngezinga lokumunca elingu-320nm.
Gwema ukulayisha ngokweqile Amapuleti e-PCR ngomkhiqizo
Njengoba kufiswa ukusebenzisa imikhiqizo eminingi ngesikhathi esisodwa, kubangela ukungcoliswa okuphambana kwamapuleti e-PCR.
Ukulayisha ngokweqile amapuleti e-PCR anemfucuza yemikhiqizo ehlukene futhi kwenza kube nzima kakhulu ukuthola amasampula.
Gcina Amarekhodi we-Aliquot PCR Reagents
Imijikelezo eqhubekayo yokuqhwaza/yokuncibilika kanye nokusetshenziswa njalo kwe-aliquot kungase konakalise ama-PCR ama-reagents, ama-enzyme kanye nama-DNTP ngokwenziwa kabusha kwekristalu.
Ngaso sonke isikhathi zama ukuqapha izinga le-aliquot elisetshenziswa ngenkathi ulungiselela amasampula azohlaziywa.
I-LIMS ekhethwayo ifaneleka kakhulu ukulawula i-inventory kanye nenani lama-reagents namasampuli afriziwe noma ancibilika.
Khetha I-Annealing Temperature Engcono Kakhulu.
Ukukhetha nokusebenzisa izinga lokushisa le-anneal elingalungile kungenye indlela imiphumela ye-PCR equkethe iphutha.
Kwesinye isikhathi, ukusabela akuhambi njengoba bekuhleliwe. Kufiswa ukunciphisa izinga lokushisa le-anealing ukuze kube lula ukusabela okuphumelelayo.
Kodwa-ke, ukwehlisa izinga lokushisa kwandisa amathuba okuba nemibono engamanga kanye nokubukeka kwe-primer dimers.
Kubalulekile ukuqinisekisa ukuhlaziya ijika elincibilikayo lapho usebenzisa amapuleti e-PCR njengoba kuyinkomba enhle yokukhetha izinga lokushisa elilungile le-anneal.
Izinsiza zesoftware yokuklama i-Primer ngokuklama, ukuhlinzekwa kwezinga lokushisa elilungile lokudonsa ngokunciphisa ngokuqondile iphutha kumapuleti e-PCR.
Ingabe Udinga Ipuleti Le-PCR Elisezingeni Eliphezulu?
Uma kwenzeka ubucabanga ukuthi ungamtholaphi umkhiqizi onokwethenjelwa weAmapuleti e-PCR. Ungabe usacinga ngoba usendaweni efanele.
Ngomusachofoza lapha ukuze uxhumane nathingemikhiqizo nesevisi yekhwalithi ephezulu ngentengo engeke iphule ibhange.
Isikhathi sokuthumela: Oct-30-2021