Inzira ya polymerase (PCR) ni bumwe muburyo buzwi bukoreshwa muri laboratoire yubuzima.
Isahani ya PCR ikorwa muri plastiki yo mucyiciro cya mbere kugirango itungwe neza kandi isesengure ingero cyangwa ibisubizo byakusanyijwe.
Bafite inkuta zoroheje kandi zihuriweho kugirango batange ihererekanyabubasha.
Mugutegura igihe nyacyo cyo gusaba, igice cyumunota wa ADN cyangwa RNA cyigunze kandi kibitswe mubisahani bya PCR.
Isahani ya PCR ikora neza mugushiraho ubushyuhe kandi igabanya umuvuduko wubushyuhe.
Nyamara, nkibikorwa byiza kandi byizewe bya plaque ya PCR, ikosa nibidasobanutse kuruhande byoroshye mugihe cyo gutunganya ingero.
Kubwibyo, niba ushishikajwe no kubona ibyiza kandi byizaIbyapa bya PCR.Nibyiza kuvugana nuwakoze plaque yizewe. Hamwe nibi wizeye ko uzabona amasezerano meza.
Hano hari ingamba zo gukurikiza kugirango wirinde kwanduza reagent cyangwa ingero no kwirinda ko ibitagenda neza byinjira mubisubizo.
Gutandukanya Ibidukikije
Ibyiza cyangwa ibibi bitabaho bitewe no kuba hari umwanda, bigatuma ushidikanya kubisubizo.
Umwanda hamwe nuwanduye bibaho muburyo butandukanye nka ADN idafitanye isano cyangwa inyongeramusaruro za chimique amaherezo bigabanya imikorere nubushobozi bwa reaction.
Hariho uburyo bwinshi bwo kugabanya cyane igipimo cyo kwanduza icyapa cya PCR.
Gukoresha akayunguruzo ka sisitemu nubundi buryo bwingirakamaro bwo kubuza umwanda kwinjira mu byitegererezo byawe unyuze mu miyoboro.
Tanga ibikoresho bisukuye rwose, bigizwe na pipeti na rake, gusa kugirango ukoreshe PCR. Ibi bizemeza kwimura byimazeyo umwanda cyangwa umwanda ukikije laboratoire.
Koresha ibibabi, Ethanol kuri pipeti, ibisate n'intebe kugirango uhanagure umwanda.
Tanga umwanya wabigenewe kuri PCR yawe yose kugirango ugabanye kwanduza uduce duto.
Koresha uturindantoki dusukuye kuri buri ntambwe hanyuma uyisimbuze kenshi.
Ibyapa bya PCR
Kugenzura Kwibanda hamwe nubuziranenge bwicyitegererezo.
Isuku yintebe nibikoresho bikoreshwa mugihe cyo gusesengura ingero hamwe na PCR bigomba gukomeza. Ni ngombwa kwemeza urwego rwubuziranenge bwintangarugero mbere yo gusesengura no gutunganya.
Mubisanzwe, abasesengura bazirikana urwego nuburinganire bwurwego rwa ADN.
Gerageza igipimo cyo kwinjiza kuri 260nm / 280nm ntigomba kuba munsi ya 1.8. Mugihe uburebure bwakurikiyeho hagati ya 230nm na 320nm bikoreshwa mukumenya umwanda.
Kurugero, umunyu wa chaotropique nibindi binyabuzima bivumburwa ku gipimo cya 230nm. Mugihe imyanda iri muri ADN ntangarugero nayo iramenyekana kandi ikagenzurwa ku gipimo cya 320nm.
Irinde kurenza urugero Isahani ya PCR hamwe nibicuruzwa
Nkuko byifuzwa gukora ibicuruzwa byinshi icyarimwe, bivamo kwanduzanya ibyapa bya PCR.
Kurenza plaque ya PCR hamwe nibicuruzwa bitandukanye byangiza imyanda kandi biragoye cyane kumenya ibyitegererezo.
Bika Inyandiko za Aliquot PCR Reagents
Gukomeza gukonjesha / kuzunguruka no gukoresha kenshi aliquot bishobora kwangiza reagent ya PCR, enzymes na DNTP binyuze muri rerystallisation.
Buri gihe ujye ugerageza gukurikirana igipimo cya aliquot ikoreshwa mugihe utegura ingero zo gusesengurwa.
LIMS ikunzwe irakwiriye kugenzura ibarura hamwe nubunini bwa reagent hamwe nicyitegererezo cyahagaritswe cyangwa cyashonze.
Tora Ubushyuhe Bwiza bwa Annealing.
Gutora no gukoresha ubushyuhe bwa annealing nubundi buryo PCR ibisubizo birimo amakosa.
Rimwe na rimwe, reaction ntigenda nkuko byateganijwe. Hifujwe kugabanya ubushyuhe bwa annealing kugirango byorohereze reaction.
Ariko, kugabanya ubushyuhe byongera amahirwe yibintu byiza na primer dimers igaragara.
Nibyingenzi kwemeza isesengura ryo gushonga umurongo mugihe ukoresheje plaque ya PCR kuko nikimenyetso cyiza cyo guhitamo ubushyuhe bukwiye bwa annealing.
Igishushanyo mbonera cya porogaramu ifasha mugushushanya, gutanga ubushyuhe bwa annealing hamwe no kugabanya amakosa yibyapa bya PCR.
Ukeneye isahani yo hejuru -Quality PCR?
Mugihe wasuzumye aho ushobora kubona uruganda rwizewe rwaIbyapa bya PCR. Ntukongere gushakisha kuko uri ahantu heza.
Mugwanezakanda hano kugirango utubwirekubicuruzwa byujuje ubuziranenge na serivisi ku giciro kitazahagarika banki.
Igihe cyo kohereza: Ukwakira-30-2021