Ukuphendula okuphumelelayo kokukhulisa, kuyadingeka ukuthi izakhi zokusabela ngakunye zikhona ekuhlolweni okulungile kulungiselelo ngalinye. Ngaphezu kwalokho, kubalulekile ukuthi kungabikho ukungcola okwenzekile.
Ikakhulu lapho ukusabela okuningi kufanele kusekwe, sekusunguliwe ukulungiselela ukuxubana okubizwa ngokuthi yi-Master ukuxubana esikhundleni se-Pipetting reagent ngayinye ngokwehlukana esitsheni ngasinye. Ukuxutshwa okulungiselelwe kwangaphambili kuyatholakala ngokuthengisa, lapho kufakwa khona izingxenye ezithile zesampula (i-Primer) namanzi. Ngenye indlela, umxube we-master ungalungiswa nguwe. Kuzo zombili lezi zinhlobonhlobo, ingxube isatshalaliswa emgodini ngamunye we-PCR ngaphandle kwethempulethi kanye nesampula ye-DNA yomuntu ngamunye ingezwa ngokwahlukana ekugcineni.
Ukusebenzisa i-Master Mix kunezinzuzo eziningana: Okokuqala, inani lezinyathelo ezizodwa zepayiphu liyancishiswa. Ngale ndlela, bobabili ubungozi bamaphutha womsebenzisi ngesikhathi sokuphambana kanye nengozi yokungcoliswa kuyancishiswa futhi, kunjalo, isikhathi siyasindiswa. Ngokuyinhloko, ukunemba kwe-Pipetting nakho kuphakeme, ngoba amavolumu amakhulu adonswa. Lokhu kulula ukukuqonda lapho ubheka idatha yezobuchwepheshe yama-Pipette: inhlamvu encane, ivolumu ephakeme, ukuphambuka kungaba. Iqiniso lokuthi wonke amalungiselelo aqhamuka kumkhumbi ofanayo unomphumela omuhle ku-homogeneity (uma kuxutshwe kahle). Lokhu futhi kuthuthukisa ukuvela kokuhlolwa.
Lapho kulungiswa i-Master Mix, okungenani ivolumu eyengeziwe engu-10% kufanele ingezwe (isib. Uma kudingeka amalungiselelo ayi-10, abale ngesisekelo se-11), ukuze ngisho nesikebhe sokugcina sigcwaliswe kahle. Ngale ndlela, (kancane) ukunganela okungalungile, kanye nomphumela wokulahleka kwesampula lapho izixazululo ezinama-dosing detergent-eziqukethe izixazululo zinganxeshezelwa. Ama-detegents aqukethe kwizixazululo ze-enzyme ezifana nama-polymerases kanye nama-master mix, abangela ukwakheka kwe-foam kanye nezinsalela ebusweni bangaphakathiAmathiphu wePipette.
Ngokuya ngohlelo nohlobo loketshezi oluzokwakhiwa, inqubo efanele ye-Pipetting (1) kufanele ikhethwe kanye nemishini efanelekile ekhethiwe. Okwezisombululo eziqukethe okokuhlanza, uhlelo oluqondile lokufuduka noma okubizwa ngokuthi "ukugcinwa okubizwa ngokuthi" amathiphu we-pipette njengenye indlela ye-popette ye-air-cushion ayanconywa. Umphumela weIthiphu ye-Ace Pipettekusekelwe endaweni ekhethekile ye-hydrophobic. I-Liquids equkethe okokuhlanza ungashiyi ifilimu eyakhayo ngaphakathi nangaphandle, ukuze ukulahlekelwa yisixazululo kungancishiswa.
Ngaphandle kokudonswa okuqondile kwazo zonke izinto, kubalulekile futhi ukuthi kungangcoliswa ukungcoliswa kwamalungiselelo. Akwanele ukusebenzisa okudliwayo kobumsulwa obuphezulu, ngoba inqubo ye-Pipetting ku-Air Cushion Pipette ingakhiqiza ama-aerosols asele ku-pipette. I-DNA engahle ifakwe ku-aerosol ingadluliselwa kusuka kusampula eyodwa kuya kwesilandelayo esiteji esilandelayo sepayipeti futhi ngaleyo ndlela kuholele ekungcolisweni. Izinhlelo zokufudumalo eziqondile ezishiwo ngenhla zinganciphisa le ngozi. Okwe-Air-Cushion Pipette Kunengqondo ukusebenzisa amathiphu wokuhlunga ukuvikela i-pipette cone ngokugcina izikhala, ama-aerosols, kanye ne-biomolecule.
Isikhathi Sokuthumela: Dec-06-2022