Ukufumana impendulo eyimpumelelo yokukhulisa, kuyimfuneko ukuba amalungu empendulo abekhona kugxininiso oluchanekileyo kulungiselelo ngalunye.Ukongeza, kubalulekile ukuba kungabikho ungcoliseko lwenzekayo.
Ngokukodwa xa uninzi lweempendulo kufuneka zisekwe, kuye kwasekwa ukulungiselela okubizwa ngokuba yi-master mix endaweni yokufaka umbhobho we-reagent nganye ngokwahlukileyo kwisitya ngasinye.Imixube elungiselelwe kwangaphambili ifumaneka ngokurhweba, apho kuphela isampula-specific components (primer) kunye namanzi zongezwa.Kungenjalo, i-master mix ingalungiswa nguwe.Kuzo zombini iinguqu, umxube usasazwa kwisitya ngasinye se-PCR ngaphandle kwetemplate kwaye isampuli ye-DNA yomntu yongezwa ngokwahlukileyo ekupheleni.
Ukusebenzisa i-master mix ineenzuzo ezininzi: Okokuqala, inani lamanyathelo okufakela umbhobho owodwa liyancipha.Ngale ndlela, zombini umngcipheko weempazamo zabasebenzisi ngexesha lokubhobhoza kunye nomngcipheko wokungcola uyancitshiswa kwaye, ngokuqinisekileyo, ixesha ligcinwa.Ngokomgaqo, ukuchaneka kombhobho kuphezulu, kuba imiqulu emikhulu ifakwe umthamo.Oku kulula ukuyiqonda xa kuhlolwa idatha yobugcisa beepipettes: Umncinci umthamo wedosi, uphezulu ukuphambuka kunokuba .Inyaniso yokuba onke amalungiselelo avela kwisitya esifanayo sinefuthe elihle kwi-homogeneity (ukuba ixutywe kakuhle).Oku kukwaphucula ukuveliswa kwakhona kwemifuniselo.
Xa ulungiselela i-master mix, ubuncinane i-10% ivolumu eyongezelelweyo kufuneka yongezwe (umz. ukuba amalungiselelo e-10 afunekayo, bala ngesiseko se-11), ukwenzela ukuba inqanawa yokugqibela izaliswe ngokufanelekileyo.Ngale ndlela, (kancinci) ukungachaneki kwemibhobho, kunye nesiphumo sokulahleka kwesampulu xa i-dosing i-detergent equlethe izisombululo zinokuhlawulwa.Iidetergents ziqulethwe kwizisombululo ze-enzyme ezifana ne-polymerases kunye ne-master mixes, ebangela ukwakheka kwamagwebu kunye neentsalela kumphezulu ongaphakathi wesiqhelo.iingcebiso zepipette.
Ngokuxhomekeke kwisicelo kunye nohlobo lolwelo oluza kuhanjiswa, indlela efanelekileyo yokufaka umbhobho (1) kufuneka ikhethwe kwaye kukhethwe izixhobo ezifanelekileyo.Kwizisombululo eziqulethe i-detergents, inkqubo yokufuduka ngokuthe ngqo okanye oko kuthiwa "ukugcinwa okuphantsi" iingcebiso ze-pipette njengenye indlela ye-air-cushion pipettes iyacetyiswa.Isiphumo seIngcebiso ye-ACE PIPETTEisekelwe kumphezulu ingakumbi hydrophobic.Ulwelo oluqulethe i-detergents alushiyi ifilimu eseleyo ngaphakathi nangaphandle, ukwenzela ukuba ukulahlekelwa kwesisombululo kuncitshiswe.
Ngaphandle kwedosi echanekileyo yawo onke amacandelo, kukwabalulekile ukuba kungabikho kungcoliseka kwamalungiselelo okwenzekayo.Akwanelanga ukusebenzisa izinto ezisetyenziswayo zokucoceka okuphezulu, kuba inkqubo yokubhobhoza kwi-pipette ye-air cushion inokuvelisa i-aerosols ehlala kwi-pipette.I-DNA enokuthi iqulethwe kwi-aerosol inokudluliselwa kwisampuli enye ukuya kwelinye kwinqanaba elilandelayo lombhobho kwaye ngaloo ndlela likhokelela ekungcoleni.Iinkqubo zokufuduswa ngokuthe ngqo ezikhankanywe ngasentla nazo zinokuwunciphisa lo mngcipheko.Kwiipayipi ze-air-cushion kunengqiqo ukusebenzisa iingcebiso zokucoca ukukhusela i-pipette cone ngokugcina i-splashes, i-aerosols, kunye ne-biomolecules.
Ixesha lokuposa: Dec-06-2022