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Nucleic Acid Extraction ke Eng?
Ka mantsoe a bonolo haholo, nucleic acid extraction ke ho tlosoa ha RNA le / kapa DNA ho tloha sampuli le tsohle tse feteletseng tse sa hlokahaleng.Ts'ebetso ea ho ntšoa e arola li-nucleic acid ho sampuli 'me e li hlahisa ka mokhoa o hlakileng oa li-eliate, tse se nang li-diluents le litšila tse ka amang lits'ebetso leha e le life tse tlase.
Lisebelisoa tsa ho Nkoa ha Nucleic Acid
Li-nucleic acid tse hloekisitsoeng li sebelisoa ka bongata ba lits'ebetso tse fapaneng, ho tloha liindastering tse fapaneng tse fapaneng.Tlhokomelo ea bophelo mohlomong ke sebaka seo e sebelisoang haholo ho sona, ka RNA e hloekisitsoeng le DNA e hlokehang bakeng sa merero e mengata e fapaneng ea tlhahlobo.
Likopo tsa ts'ebetso ea nucleic acid ho tsa bophelo bo botle li kenyelletsa:
- Next Generation Sequencing (NGS)
- SNP Genotyping e thehiloeng ho Amplification
- Array-based Genotyping
- Thibelo tšilo ea enzyme
- E sekaseka ka ho sebelisa li-Enzymes tse Fetohang (mohlala, Ligation le Cloning)
Ho boetse ho na le likarolo tse ling tse fetang tlhokomelo ea bophelo moo ho sebelisoang nucleic acid, ho kenyeletsoa empa ho sa felle feela ho tlhahlobo ea bontate, forensics le genomics.
Histori e Khutšoanyane ea Nōko ea Nucleic Acid
Ho ntšoa ha DNAe qalile khale haholo, ka ho itšehla thajana ha pele ho entsoe ke ngaka ea Switzerland e bitsoang Friedrich Miescher ka 1869. Miescher o ne a tšepile ho rarolla melao-motheo ea bophelo ka ho fumana hore na lik'hemik'hale li na le lisele tse kae.Ka mor'a ho hlōleha ho sebelisa li-lymphocyte, o ile a khona ho fumana mokelikeli o sa tloaelehang oa DNA ho tsoa ho li-leukocyte tse fumanoang pus holim'a li-bandage tse lahliloeng.O entse sena ka ho eketsa asiti ebe alkali seleng ho tloha cytoplasm ea sele, ebe o theha protocol ea ho arola DNA ho liprotheine tse ling.
Ka mor'a lipatlisiso tsa motheo tsa Miescher, bo-rasaense ba bang ba bangata ba tsoetse pele le ho hlahisa mekhoa ea ho arola le ho hloekisa DNA.Edwin Joseph Cohn, rasaense oa protheine o ile a hlahisa mekhoa e mengata ea ho hloekisa protheine nakong ea WW2.O ne a ikarabella bakeng sa ho arola karoloana ea serum albumin ea plasma ea mali, e leng ea bohlokoa ho boloka khatello ea osmotic methapong ea mali.Sena se ne se le bohlokoa bakeng sa ho boloka masole a phela.
Ka 1953 Francis Crick, hammoho le Rosalind Franklin le James Watson, ba ile ba etsa qeto ea sebōpeho sa DNA, ho bontša hore e entsoe ka liketane tse peli tse telele tsa nucleic acid nucleotide.Katleho ena e sibollotsoeng e ile ea betlela Meselson le Stahl tsela, ba ileng ba khona ho etsa protocol ea density gradient centrifugation ho arola DNA ho libaktheria tsa E. Coli ha ba ntse ba bonts'a phetisetso ea semi-conservative ea DNA nakong ea liteko tsa bona tsa 1958.
Mekhoa ea ho Ntša Nucleic Acid
Mehato e 4 ea ho ntšoa ha DNA ke efe?
Mekhoa eohle ea ho hula e itšetlehile ka mehato e tšoanang ea motheo.
Tšitiso ea Lisele.Mokhahlelo ona, oo hape o tsejoang e le cell lysis, o kenyelletsa ho pshatla lerako la sele le/kapa lera la sele, e le hore ho lokolloe maro a intra-cellular a nang le li-nucleic acid tse khahlang.
Ho Tlosa Litšila Tse sa Batloeng.Sena se kenyelletsa membrane lipids, liprotheine le li-nucleic acid tse ling tse sa batleheng tse ka sitisang lits'ebetso tse tlase.
Ho behwa ka thoko.Ho na le mekhoa e mengata e fapaneng ea ho arola li-nucleic acid tse khahlang ho lysate e hloekisitsoeng eo u e entseng, e oelang lipakeng tsa mekhahlelo e 'meli e meholo: boemo ba tharollo kapa bo tiileng (sheba karolo e latelang).
Ho tsepamisa maikutlo.Ka mor'a hore li-nucleic acid li arohane le litšila tse ling tsohle le li-diluents, li hlahisoa ka mokhoa o hlakileng haholo.
Mefuta e 'meli ea ho lema
Ho na le mefuta e 'meli ea tlhahiso ea asiti ea nucleic - mekhoa e thehiloeng ho tharollo le mekhoa ea boemo bo tiileng.Mokhoa o thehiloeng ho tharollo o boetse o tsejoa e le mokhoa oa ho ntša lik'hemik'hale, kaha o kenyelletsa ho sebelisa lik'hemik'hale ho senya sele le ho fumana lisebelisoa tsa nucleic.Sena e ka ba ho sebelisa metsoako ea lintho tse phelang tse kang phenol le chloroform, kapa metsoako e seng kotsi haholo 'me ka hona e khothaletsoa haholoanyane joalo ka Proteinase K kapa silika gel.
Mehlala ea mekhoa e fapaneng ea ho ntša lik'hemik'hale ho senya sele e kenyelletsa:
- Ho phatloha ha lera la Osmotic
- Tšilo ea tšilo ea 'mele ea lebota la lisele
- Solubilisation ea lera
- Ka li-detergents
- Ka phekolo ea alkali
Mekhoa e tiileng ea mmuso, eo hape e tsejoang e le mekhoa ea mochini, e kenyelletsa ho sebelisa hampe tsela eo DNA e sebelisanang ka eona le substrate e tiileng.Ka ho khetha sefaha kapa molek'hule eo DNA e tla tlama ho eona empa mohlahlobi a ke ke a etsa joalo, hoa khoneha ho arola tse peli.Mehlala ea mekhoa e tiileng ea ho ntša likhahla ho kenyelletsa ho sebelisa silica le lifaha tsa makenete.
Ho Hlalosoa ha Lifaha Tsa Magnetic
Mokhoa oa ho Ntša Lifaha Tsa Magnetic
Monyetla oa ho ntšoa ka lifaha tsa makenete o ile oa hlokomeloa ka lekhetlo la pele ho patent ea US e kentsoeng ke Trevor Hawkins, bakeng sa setsi sa lipatlisiso sa Whitehead Institute.Patent ena e ile ea lumela hore ho ne ho ka khoneha ho ntša liphatsa tsa lefutso ka ho li tlama ho sesebelisoa se tiileng sa tšehetso, seo e ka bang sefaha sa makenete.Molao-motheo ke hore u sebelise sefaha sa makenete se sebetsang hantle haholo seo liphatsa tsa lefutso li tla itlama ho sona, ebe joale se ka aroloa ho se matla ka ho sebelisa matla a khoheli ka ntle ho sekepe se tšoereng sampole.
Ke Hobane'ng ha U Sebelisa Magnetic Bead Extraction?
Theknoloji ea ho ntša lifaha tsa Magnetic e ntse e ata haholo, ka lebaka la bokhoni boo e nang le bona bakeng sa mekhoa e potlakileng le e sebetsang ea ho ntša.Haufinyane tjena ho bile le lintlafatso tsa lifaha tsa makenete tse sebetsang hantle haholo tse nang le li-buffer system tse loketseng, tse entseng hore ho khonehe ho ntša nucleic acid le mokhoa oa ho sebetsa o bobebe haholo oa lisebelisoa ebile o baballa chelete e ngata.Hape, mekhoa ea ho ntša lifaha tsa makenete ha e kenyelle mehato ea centrifugation e ka bakang matla a ho kuta a senyang likarolo tse telele tsa DNA.Sena se bolela hore likhoele tse telele tsa DNA li lula li tiile, e leng ntho ea bohlokoa litekong tsa genomics.
Nako ea poso: Nov-25-2022