Ibintu byo kwivanga muri PCR reaction

Mugihe cya PCR, ibintu bimwe na bimwe bibangamiwe.
Bitewe nubunini bwa PCR, kwanduza bifatwa nkimwe mubintu byingenzi bigira ingaruka kubisubizo bya PCR kandi birashobora kubyara ibisubizo byiza.
Kunegura kimwe nisoko zitandukanye ziganisha kubisubizo bibi. Niba kimwe cyangwa byinshi byingenzi bya PCR kuvanga cyangwa reaction yongerewe ubwabyo byabujijwe cyangwa bibangamiwe, isuzuma ribi rishobora kubangamirwa. Ibi birashobora kuganisha ku kugabanya imikorere ndetse nibisubizo bibi byo mubi.
Usibye kubuza, gutakaza intego ya acide nucleic irashobora kubaho kubera kohereza no / cyangwa imiterere mbere yo gutegura icyitegererezo. By'umwihariko, ubushyuhe bwo hejuru cyangwa ububiko budahagije bishobora gutera kwangirika kwa selile na acide ya nucleic. Akagari ka selile na tissue na paraffin bazwiho bitewe no guhagarika umutima hamwe nikibazo gihoraho (reba Imibare 1 na 2). Muri ibi bihe, ndetse no kwigunga neza no kweza ntibizafasha.

Igishushanyo 1 | Ingaruka zo Kwishura Kubunyangamugayo bwa ADN
Agarose Gel Electrophores yerekanye ko ubwiza bwa ADN bwitaruye muminota ya Paraffin yatunganijwe neza. ADN yinshuro zisanzwe zo gutandukana zari zihari mubyo bitewe nuburyo bwo gukosora. ADN yabitswe gusa mugihe yashyizwe mu byitegererezo kavukire kavukire no muri rubisiyo itabogamye. Gukoresha acide bifatika bikosora cyangwa bidasubirwaho, acide ya acide ikubiyemo acide yavuyemo igihombo gikomeye cya ADN. Igice gisigaye kiracitsemo ibice cyane.
Ibumoso, uburebure bwibice bigaragarira mu kilobase ibice (KBP)

Igishushanyo 2 | Gutakaza ubusugire bwintego za acide nucleic
. Synthesis ya ADN izakomeza kugaragara ku gice gito. Ariko, niba urubuga rwambere rwa primer rwabuze kumurongo wa ADN, gusa gusa umurongo wo kumurongo bibaho. Mu rubanza rwiza cyane, ibice birashobora kugarura, ariko umusaruro uzaba muto kandi uri munsi y'urwego rwo gutahura.
. Mugihe cyicyiciro gishyushye cyane, primers izashonga kure ya matrix ADN kandi ntibuzana no mubihe bitarangwamo.
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Ikindi kibazo gikunze kugaragara muri diagnostique ya molecular nicyo kitari cyiza-cyiza acide ya aside iricleic ugereranije nibikuramo chenol-chloroform. Mubihe bikabije, ibi birashobora kubahirizwa nibibi. Igihe kinini gishobora gukizwa muguteka Lyailing cyangwa izogo zo mu bwoko bwa selile, ariko ubu buryo akenshi buturuka kuri PCR nkeya kubera kurekurwa kwa nucleic bihagije.

Irwanya ibikorwa bya Polymeyese mugihe cyongerera

Muri rusange, ihagarikwa rikoreshwa nk'igitekerezo cyo gusobanura ibintu byose biganisha ku bisubizo bya PCR. Muburyo bukomeye bwibinyabuzima, ihagarikwa rigarukira gusa mubikorwa bya enzyme, ni ukuvuga bigabanya cyangwa birinda guhindura ibicuruzwa hamwe nurubuga rwa ADN cyangwa Formactor yarwo (urugero: GG2 + kuri Taq ADN Polymeyese).
Ibigize murugero cyangwa bukurya bitandukanye birimo reagents birashobora kubuza enzyme cyangwa gutesha agaciro abakora amashuri cyangwa ngo badakora polymerise kugirango bagabanye cyangwa ibinyoma bya PCR ibiva.
Nyamara, imikoranire myinshi hagati yibigize reaction hamwe nintego-irimo acide ya nucleic nayo yagenwe nka 'PCR ibibuza'. Ubusugire bw'akagari bumaze guhungabana mu bwigunge no kwigunga no gucikamo ibirafu birasohoka, imikoranire hagati y'icyitegererezo n'icyiciro cyayo kiboneka hamwe n'icyiciro gikomeye gishobora kubaho. Kurugero, 'Scavengers' irashobora guhambira ADN imwe- cyangwa ibiri yashinze imikoranire idasanzwe kandi ikabangamira kwigunga no kweza kugabanya umubare wintego amaherezo ugera kuri icyombo cya PCR kigera kuri posel.
Muri rusange, abagizi ba nabi ba PCR bahari mumazi menshi kandi bakoresheje ibizamini byo gusuzuma ubuvuzi (Urea mu nkari.

Abagizi ba nabi ba PCR bakomeye barashobora kuboneka muri bagiteri na eukaryotic selile, idahwitse ADNA, Guhuza Macronolecules yimibare ya tissue nibikoresho bya laboratoire nka gants. Gusukura acide ya nucleic mugihe cyangwa nyuma yo gukuramo nuburyo bukunzwe bwo gukuraho abarashi PCR.
Uyu munsi, ibikoresho bitandukanye byo gukuramo byikora birashobora gusimbuza protocole nyinshi, ariko 100% kugarura no / cyangwa kwezwa intego ntabwo bigeze kugerwaho. Ibishobora kubamo birashobora gukomeza kuba muri acide nucleic yasukuye cyangwa bishobora kuba byarateganijwe. Ingamba zitandukanye zihari kugirango ugabanye ingaruka z'abagizi ba. Guhitamo Polymeyese bikwiye birashobora kugira ingaruka zikomeye kubikorwa bya MIBIET. Ubundi buryo bwemejwe kugirango kugabanya imuga pcr biroroshye kwibanda cyane cyangwa gusaba inyongera nka BSA.
Kubuza Ibisubizo bya PCR birashobora kugaragazwa no gukoresha inzira yimbere (IPC).
Kwitaho bigomba gukuraho reagents zose nibindi bisubizo mubikoresho byo gukuramo, nka Edta, Cetab, SDS, Isopropanol na acide, muri acide ya nucleic yo gukaraba. Ukurikije kwibanda kwabo, barashobora gukora cyangwa kubuza PCR.