Ibipimo ngenderwaho bikubiyemo uburyo bwiza bwogutezimbere no gushiraho no guhuza, kwemerera gukora igihe kirekire cyiza - kidashingiye kubakoresha. Ibipimo ngenderwaho byemeza ibisubizo byujuje ubuziranenge, kimwe no kubyara no kugereranya.
Intego ya (classique) PCR nigisekuru cyibisubizo byizewe kandi byororoka. Kuri Porogaramu zimwe, umusaruro waIbicuruzwa bya PCRni ngombwa. Kuri ibyo bitekerezo, hagomba kwitonderwa kugirango ingero zidahungabana kandi ko PCR ikora neza. By'umwihariko, ibi bisobanura kugabanya itangizwa ryanduye rishobora kuganisha kubisubizo byiza cyangwa ibinyoma bibi cyangwa bikabuza PCR kwitwara. Ikigeretse kuri ibyo, imiterere yimyitwarire igomba kuba imwe ishoboka kuri buri cyitegererezo cyumuntu kugikora kandi ikanimurirwa mubitekerezo byakurikiyeho (muburyo bumwe). Ibi bivuga ibigize reaction kimwe n'ubwoko bwo kugenzura ubushyuhe muri cycler. Amakosa y'abakoresha, birumvikana ko agomba kwirindwa bishoboka.
Hasi, tuzerekana imbogamizi zihura nazo mugihe cyo kwitegura no mugihe cyose cya PCR - hamwe nuburyo bwo gukemura ibibazo bibaho kubijyanye nibikoresho nibikoresho bikoreshwa muguhuza ibikorwa bya PCR.
Gutegura ibisubizo
Ikwirakwizwa ryibigize reaction muri PCR-yamato, cyangwa amasahani, bikubiyemo ibibazo byinshi bigomba kuneshwa:
Ibisabwa
Igipimo nyacyo kandi cyuzuye cyibigize buriwese ntigisabwa mugihe ugamije ibintu bisa nkibishoboka. Usibye tekinike nziza yo kuvoma, guhitamo igikoresho cyiza nibyingenzi. PCR master-ivanga kenshi irimo ibintu byongera ubwiza cyangwa bikabyara ifuro. Mugihe cyo kuvoma, ibyo biganisha kumazi menshi yainama, bityo kugabanya imiyoboro nyayo. Gukoresha sisitemu yo gutanga mu buryo butaziguye cyangwa ubundi buryo bwa pipette butagereranywa no guhanagura birashobora kunonosora neza kandi neza neza inzira yo kuvoma.
Umwanda
Mugihe cyo gutanga, hakorwa aerosole, iyo yemerewe kugera imbere muri pipeti, irashobora kwanduza urundi rugero mugihe gikurikira. Ibi birashobora gukumirwa ukoresheje akayunguruzo cyangwa sisitemu yo kwimura abantu.
Ibikoreshwa nkainama, inzabya n'amasahani bikoreshwa mubikorwa bya PCR ntibigomba kuba birimo ibintu bibangamira icyitegererezo cyangwa kubeshya ibisubizo. Harimo ADN, DNase, RNase na PCR inhibitor, hamwe nibice bishobora kuva mubintu mugihe cyo kubyitwaramo - ibintu bizwi nka leachables.
Ikosa ryabakoresha
Ingero nyinshi zitunganijwe, niko ibyago byo kwibeshya. Birashobora kubaho byoroshye ko icyitegererezo cyinjijwe mu cyombo kitari cyo cyangwa iriba ribi. Izi ngaruka zirashobora kugabanuka cyane mukumenyekanisha byoroshye amariba. Binyuze muri automatisation yo gutanga intambwe, "ibintu byabantu", ni ukuvuga amakosa hamwe nuburyo butandukanye bujyanye n’abakoresha, bigabanywa, bityo bikongera imyororokere, cyane cyane mubibazo bito bito. Ibi bisaba amasahani yumutekano uhagije kugirango ukoreshwe kumurimo. Kode ya barcode itanga imashini isomeka, yoroshya icyitegererezo mugikorwa cyose.
Porogaramu ya thermocycler
Gutegura igikoresho birashobora kwerekana ko bitwara igihe kimwe nibishobora kwibeshya. Ibintu bitandukanye bya PCR yubushyuhe bwa cycler birakorana kugirango byoroshe iyi ntambwe kandi cyane cyane, kugirango itekane:
Igikorwa cyoroshye hamwe nubuyobozi bwiza bukoreshwa nishingiro rya progaramu nziza. Kubaka kuriyi fondasiyo, ijambo ryibanga ririnzwe ukoresha bizarinda gahunda zumuntu guhindurwa nabandi bakoresha. Niba abanyamagare benshi (b'ubwoko bumwe) barimo gukoreshwa, nibyiza niba porogaramu ishobora kwimurwa biturutse ku gikoresho kimwe ikajya mu kindi binyuze kuri USB cyangwa guhuza. Porogaramu ya mudasobwa ituma ubuyobozi bukuru kandi bwizewe bwa porogaramu, uburenganzira bwabakoresha ninyandiko kuri mudasobwa.
PCR ikora
Mugihe cyo kwiruka, ADN yongerewe imbaraga mubwato bwitwara, aho buri cyitegererezo kigomba gukorerwa ibintu bimwe, bihoraho. Ibice bikurikira birakenewe mubikorwa:
Kugenzura ubushyuhe
Ubwiza buhebuje mukugenzura ubushyuhe hamwe nuburinganire bwumukino wamagare niyo shingiro ryubushyuhe bwubushakashatsi bwintangarugero zose. Ubwiza buhanitse bwibintu byo gushyushya no gukonjesha (peltier element), hamwe nuburyo ibyo bihuza na blok, nimpamvu zifatika zizagaragaza ingaruka ziterwa nubushyuhe buzwi nka "inkurikizi"
Umwuka
Ubwinshi bwibice bigize reaction ya buriwese ntibigomba guhinduka mugihe cyibisubizo biterwa no guhumeka. Bitabaye ibyo, birashoboka ko ari bike cyaneIbicuruzwa bya PCRirashobora kubyara, cyangwa ntayo rwose. Ni ngombwa rero kugabanya imyuka ihumeka neza. Muri iki gihe, umupfundikizo ushyushye wa thermocycler hamwe na kashe yubwato bikora mu ntoki. Uburyo butandukanye bwo gufunga burahari kuriIsahani ya PCR (ihuza: Ikidodo), aho kashe nziza igerwaho hifashishijwe gufunga ubushyuhe. Ibindi bifunga nabyo birashobora kuba byiza, mugihe cyose igitutu cyo guhuza umupfundikizo wamagare gishobora guhinduka kashe yatoranijwe.
Ibipimo ngenderwaho birahari murwego rwo kurinda ibisubizo nyabyo kandi byororoka mugihe kirekire. Ibi bikubiyemo gufata neza ibikoresho kugirango tumenye ko buri gihe bimeze neza. Ibikoreshwa byose bigomba kuba byujuje ubuziranenge murwego rwinshi rwakozwe, kandi kuboneka kwizewe bigomba kuba byemewe.
Igihe cyoherejwe: Ugushyingo-29-2022