Ke Eng e Lokelang ho Nahana ka Pipeting PCR Mixtures?

Bakeng sa karabelo e atlehileng ea amplification, hoa hlokahala hore likarolo tsa karabelo ka bomong li be teng ka har'a mahloriso a nepahetseng tokisong ka 'ngoe. Ho phaella moo, ke habohlokoa hore ho se be le tšilafalo e hlahang.

Haholo-holo ha mekhoa e mengata e tlameha ho hlophisoa, e thehiloe ho lokisa seo ho thoeng ke motsoako o moholo ho e-na le ho kenya liphaephe tse fapaneng ka har'a sekepe ka seng. Metsoako e lokiselitsoeng esale pele e fumaneha khoebong, eo ho eona ho kenyelletsoang likarolo tse khethehileng tsa sampole (primer) le metsi. Ntle le moo, motsoako oa master o ka lokisoa ke uena. Ka mefuta e 'meli, motsoako o ajoa sekepeng se seng le se seng sa PCR ntle le template 'me sampole ea DNA ka bomong e eketsoa ka thoko qetellong.

Ho sebelisa motsoako o moholo ho na le melemo e 'maloa: Ntlha ea pele, palo ea mehato e le' ngoe ea pipetting e fokotsehile. Ka tsela ena, bobeli ba kotsi ea liphoso tsa basebelisi nakong ea pipetting le kotsi ea tšilafalo e fokotsehile 'me, ha e le hantle, nako e bolokiloe. Ha e le hantle, ho nepahala ha pipetting ho boetse ho phahame, kaha li-volume tse kholo li fokotsoa. Sena se bonolo ho se utloisisa ha u hlahloba lintlha tsa tekheniki tsa li-pipette: Ha molumo o fokotsehile, liphapang li ka ba tse phahameng. Taba ea hore litokisetso tsohle li tsoa sekepeng se le seng se na le phello e ntle ho homogeneity (haeba e kopane hantle). Sena se boetse se ntlafatsa ho ikatisa ha liteko.

Ha u lokisetsa motsoako o mongata, bonyane 10% ea molumo o eketsehileng o lokela ho eketsoa (mohlala, haeba ho hlokahala litokisetso tse 10, bala ho latela 11), e le hore esita le sekepe sa ho qetela se tlale hantle. Ka tsela ena, (hanyenyane) ho se nepahale ha lipeipi, le phello ea tahlehelo ea sampole ha litharollo tse nang le li-detergent li ka lefshoa. Li-detergents li fumaneha ka har'a litharollo tsa enzyme tse kang polymerase le li-mix mixes, tse bakang foam le masala ka har'a bokaholimo ba lintho tse tloaelehileng.malebela a pipette.

Ho ipapisitsoe le ts'ebeliso le mofuta oa mokelikeli o tla fanoa, ho lokela ho khethoa mokhoa o nepahetseng oa lipeipi (1) 'me ho khethoe lisebelisoa tse nepahetseng. Bakeng sa litharollo tse nang le li-detergents, tsamaiso e tobileng ea ho falla kapa seo ho thoeng ke "ho boloka tlaase" malebela a pipette e le mokhoa o mong oa li-pipette tsa moea oa moea. Phello eaKeletso ea ACE PIPETTEe thehiloe holim'a sebaka se khethehileng sa hydrophobic. Mekelikeli e nang le li-detergents ha e tlohele filimi e setseng ka hare le ka ntle, e le hore tahlehelo ea tharollo e fokotsehe.

Ntle le dosing e nepahetseng ea likarolo tsohle, ke habohlokoa hape hore ho se be le tšilafalo ea litokisetso e hlahang. Ha hoa lekana ho sebelisa lisebelisoa tsa bohloeki bo phahameng, hobane mokhoa oa pipetting ka pipette ea moea o ka hlahisa li-aerosol tse setseng ka pipette. DNA e ka 'nang ea e-ba teng ka har'a aerosol e ka fetisetsoa ho tloha sampuling e' ngoe ho ea ho e 'ngoe ka mohato o latelang oa pipetting' me kahoo ea lebisa tšilafalo. Litsamaiso tse tobileng tsa phalliso tse boletsoeng ka holimo le tsona li ka fokotsa kotsi ena. Bakeng sa li-pipette tsa moea-cushion hoa utloahala ho sebelisa malebela a sefa ho sireletsa khoune ea pipette ka ho boloka li-splashes, li-aerosols le li-biomolecules.


Nako ea poso: Dec-06-2022