Kugirango bigende neza kuri amplification reaction, birakenewe ko buri reaction yibikorwa igaragara muburyo bwiza muri buri myiteguro. Byongeye kandi, ni ngombwa ko hatabaho umwanda.
Cyane cyane mugihe hagomba gushyirwaho ibisubizo byinshi, hashyizweho gutegura icyitwa mix mix aho guhinduranya buri reagent ukwayo muri buri cyombo. Imvange yabanje gushyirwaho iraboneka mubucuruzi, aho byongeweho gusa icyitegererezo cyihariye (primer) n'amazi byongeweho. Ubundi, master mix irashobora gutegurwa wenyine. Muri ubwo buryo bwombi, imvange ikwirakwizwa kuri buri cyombo cya PCR nta shusho kandi icyitegererezo cya ADN cyongeweho ukwacyo kurangiza.
Gukoresha master mix ifite ibyiza byinshi: Icya mbere, umubare wintambwe imwe yo kugabanuka uragabanuka. Muri ubu buryo, ibyago byombi byamakosa yabakoresha mugihe cyo gutembera hamwe ningaruka zo kwandura bigabanutse kandi, byanze bikunze, igihe cyarazigamiwe. Ihame, imiyoboro ihanitse nayo iri hejuru, kubera ko ingano nini zafashwe. Ibi biroroshye kubyumva mugihe ugenzura amakuru ya tekiniki ya pipettes: Nubunini buke bwa dosiye, niko gutandukana bishobora kuba. Kuba imyiteguro yose iva mu cyombo kimwe igira ingaruka nziza ku bahuje ibitsina (niba bivanze neza). Ibi kandi bitezimbere imyororokere yubushakashatsi.
Mugihe utegura master mix, byibuze 10% byongeweho bigomba kongerwaho (urugero niba hakenewe imyiteguro 10, ubare ukurikije 11), kuburyo nubwato bwa nyuma bwuzuye neza. Muri ubu buryo, (bito) imiyoboro idahwitse, hamwe ningaruka zo gutakaza icyitegererezo mugihe cyo gukuramo imiti irimo ibikoresho bishobora kwishyurwa. Imyanda ikubiye mubisubizo bya enzyme nka polymerase hamwe no kuvanga master, bigatera kwibumbira hamwe nibisigara hejuru yimbere mubisanzweinama.
Ukurikije porogaramu n'ubwoko bw'amazi agomba gutangwa, tekinike nziza yo kuvoma (1) igomba guhitamo nibikoresho byatoranijwe. Kubisubizo birimo ibikoresho byogajuru, sisitemu yo kwimura mu buryo butaziguye cyangwa inama yiswe "kugabanuka gake" nk'uburyo bwo guhinduranya imiyoboro yo mu kirere irasabwa. Ingaruka yaACE PIPETTE inamaishingiye ku buso bwa hydrophobique. Amazi arimo ibikoresho byo kwisiga ntabwo asiga firime isigaye imbere no hanze, kugirango igihombo kibe gito.
Usibye gukuramo neza ibice byose, ni ngombwa kandi ko hatabaho kwanduza imyiteguro. Ntabwo bihagije gukoresha ibikoreshwa byera cyane, kuko inzira yo gutobora mumashanyarazi yo mu kirere irashobora kubyara aerosole iguma muri pipeti. ADN ishobora kuba iri muri aerosol irashobora kwimurwa ikava murugero rumwe ikajya kurindi murwego rukurikira rwo kuvoma bityo bigatera kwanduza. Sisitemu yo kwimura itaziguye yavuzwe haruguru irashobora kandi kugabanya ibi byago. Kubijyanye na air-cushion pipettes birumvikana gukoresha inama zo kuyungurura kugirango urinde umuyoboro wa pipette ugumana ibice, aerosole, na biomolecules.
Igihe cyo kohereza: Ukuboza-06-2022