ⅠNasiti ea ucleiciselelekela
Nucleic acid e arotsoe ka deoxyribonucleic acid (DNA) le ribonucleic acid (RNA), eo RNA e arotsoeng ka ribosomal RNA(rRNA), lenģosa RNA(mRNA) le ho fetisetsa RNA(tRNA) ho latela mesebetsi ea eona e fapaneng.DNA e tsepamisitsoe haholo khubung, mitochondria le chloroform, ha RNA e ajoa haholo ho cytoplasm.E le motheo oa liphatsa tsa lefutso, ho ntšoa ha nucleic acid ho bapala karolo ea bohlokoa haholo lipatlisisong tsa baeloji ea limolek'hule le tlhahlobo ea limolek'hule tsa bongaka.Ho tsepamisa mohopolo le ho hloeka ha nucleic acid e nkiloeng ho tla ama ka ho toba PCR e latelang, tatellano, kaho ea vector, tšilo ea enzyme le liteko tse ling.
Ⅱ Mokhoa oa ho ntša asiti ea nucleic le mokhoa oa ho hloekisa
① Mokhoa oa ho ntša phenol/chloroform
Phenol/chloroform extraction ke mokhoa oa khale oa ho ntšoa ha DNA, oo haholo-holo o sebelisang metsoako e 'meli e fapaneng ea manyolo ho phekola lisampole, ho qhala DNA e thehiloeng ho nucleic acid mothating oa metsi, lipids karolong ea manyolo, le liprotheine lipakeng tsa mekhahlelo e 'meli.Mokhoa ona o na le melemo ea theko e tlaase, bohloeki bo phahameng le phello e ntle.Melemo ke ts'ebetso e rarahaneng le nako e telele.
② Mokhoa oa Trizol
Mokhoa oa Trizol ke mokhoa oa khale oa ho ntša RNA.Mokhoa oa Trizol o arotsoe ka karolo ea metsi le karolo ea manyolo ka mor'a hore centrifugation e be le chloroform, eo RNA e qhibilihisang ka eona karolong ea metsi, karolo ea metsi e fetisetsoa ho tube e ncha ea EP, pula e fumanoa ka mor'a ho eketsa isopropanol, ebe ho hloekisoa ha ethanol.Mokhoa ona o loketse ho ntšoa ha RNA ho tsoa ho lisele tsa liphoofolo, lisele le libaktheria.
③ Mokhoa oa tlhoekiso ea kholomo ea Centrifugal
Centrifuge kholomo tlhoekiso mokhoa ka adsorb DNA ka ho khetheha ka silicon matrix adsorption thepa e khethehileng, ha RNA le protheine ka feta ka thelelo, 'me joale sebelisa letsoai le phahameng tlaase PH ho kopanya nucleic acid, tlaase letsoai phahameng PH bohlokoa elution ho arola le ho hloekisa acid e nucleic.Melemo ke khatello e phahameng ea tlhoekiso, botsitso bo phahameng, ha ho na tlhoko ea solvent ea manyolo, le litšenyehelo tse tlase.Bothata ke hore e hloka ho centrifuged mohato ka mohato, mehato e mengata ea ts'ebetso.
④ Mokhoa oa lifaha tsa makenete
Mokhoa oa "magnetic beads" ke ho arola sampole ea lisele tsa lisele ka har'a lysate, ho lokolla acid ea nucleic ka har'a sampole, ebe limolek'hule tsa nucleic acid li ts'oaroa ka ho khetheha holim'a sefaha sa makenete, ha litšila tse kang liprotheine le tsoekere li ntse li le teng. mokelikeli.Ka mehato ea ho petsoha ha lisele tsa lisele, ho tlama lifaha tsa makenete ka nucleic acid, ho hlatsuoa ha nucleic acid, nucleic acid elution, joalo-joalo, qetellong ho fumanoe asiti e hloekileng ea nucleic.Melemo ke ts'ebetso e bonolo le ts'ebeliso ea nako e khuts'oane, ntle le tlhoko ea mohato oa centrifugation.E na le litlhoko tse tlase tsa tekheniki mme e khona ho etsa ts'ebetso ea othomathike le ea bongata.Motsoako o khethehileng oa magnetic bead le nucleic acid o etsa hore nucleic acid e ntšitsoe ka mahloriso le bohloeki bo phahameng.Bothata ke hore theko ea hona joale ea 'maraka e batla e le theko e boima.
⑤ Mekhoa e meng
Ho phaella mekhoeng e mene e ka holimo, ho na le ho qhekella ho belang, mokhoa oa letsoai o tsitsitseng, mokhoa oa anionic detergent, mokhoa oa ultrasonic le mokhoa oa enzymatic, joalo-joalo.
Ⅲ Mofuta oa ho ntšoa ha nucleic acid
Foregene e na le sethala sa Direct PCR se etelletseng pele lefatšeng, sethala sa likholomo tse peli tsa RNA Isolation (DNA-feela + RNA feela ).Lihlahisoa tsa mantlha li kenyelletsa lisebelisoa tsa ho itšehla thajana tsa DNA/RNA, PCR le Direct PCR reagents letoto la li-molecular lab reagents.
① Kakaretso ea RNA e nkiloeng
Kakaretso ea lisampole tsa RNA tse nkiloeng li kenyelletsa mali, lisele, lisele tsa liphoofolo, limela, likokoana-hloko, joalo-joalo. Bohloeki bo phahameng le khatello e phahameng ea RNA kaofela e ka fumanoa ka ho ntša RNA e feletseng, e ka sebelisoang ho RT-PCR, tlhahlobo ea chip, phetolelo ea in vitro, cloning ea molek'hule, Dot Blot le liteko tse ling.
Foregene amanangRNA Isolation Kits
Animal Total RNA Isolation Kit--Ka potlako le ka mokhoa o nepahetseng ntša RNA ea boleng bo holimo le ea boleng bo holimo ho tsoa liseleng tse fapaneng tsa liphoofolo.
Cell Total RNA Isolation Kit--RNA e hloekisitsoeng haholo le ea boleng bo holimo e ka fumanoa liseleng tse fapaneng tsa setso ka metsotso e 11.
Semela Kakaretso ea RNA Isolation Kit--Ka potlako ntša RNA ea boleng bo holimo ho lisampole tsa semela tse nang le polysaccharide e tlase le polyphenol content.
Viral RNA Isolation Kit--Arola ka potlako le ho hloekisa RNA ea kokoana-hloko ho lisampole tse kang plasma, serum, maro a 'mele a se nang lisele le li-supernatant tsa cell culture.
② Ho ntšoa ha DNA ea Genomic
Mehlala ea ho ntšoa ha DNA ea Genomic e kenyelletsa mobu, mantle, mali, lisele, lisele tsa liphoofolo, limela, likokoana-hloko, joalo-joalo. Genomic DNA extraction e ka sebelisoa ka tšilo ea lijo ea enzyme, kaho ea laebrari ea DNA, PCR, ho lokisetsa li-antibody, Western blot hybridization analysis, gene chip, phahameng. - tatellano ea litaba le liteko tse ling.
Foregene amanangDNA Isolation Kits
DNA ea Lithane tsa Phoofolo Isolation Kit- Ho ntšoa ka potlako le tlhoekiso ea DNA ea genomic ho tsoa mehloling e mengata, joalo ka lisele tsa liphoofolo, lisele, joalo-joalo.
Mali a DNA Midi Kit (1-5ml)--Ka potlako hloekisa DNA ea boleng bo holimo ho tsoa mali a anticoagulated (1-5ml).
Buccal Swab / FTA Card DNA Isolation Kit--Ka potlako hloekisa DNA ea boleng bo holimo ho tsoa lisampoleng tsa buccal swab/FTA Card.
Semela sa DNA Isolation Kit--Hloekisa kapele 'me u fumane DNA ea boleng bo holimo ho tsoa lisampoleng tsa semela (ho kenyeletsoa lisampole tsa polysaccharides le polyphenol limela)
③ Ho ntšoa ha polasetiki
Plasmid ke mofuta oa selikalikoe sa molek'hule oa DNA liseleng, e leng sejari se tloaelehileng sa ho kopanya DNA.Mokhoa oa ho ntša plasmid ke ho tlosa RNA, ho arola plasmid ho bacterial genomic DNA, le ho tlosa protheine le litšila tse ling ho fumana plasmid e batlang e hloekile.
Kakaretso ea Plasmid Mini Kit--Ka potlako hloekisa plasmid DNA ea boleng bo holimo ho libaktheria tse fetotsoeng bakeng sa liteko tse tloaelehileng tsa baeloji ea limolek'hule joalo ka phetoho le tšilo ea enzyme.
④ Mefuta e meng ea ho ntša, miRNA e ntšoa, joalo-joalo.
Animal miRNA Isolation Kit- Ka potlako le ka mokhoa o atlehileng o ntša likhechana tsa RNA tsa 20-200nt miRNA, siRNA, snRNA ho tsoa liseleng tse fapaneng tsa liphoofolo.
Ⅳ Litlhokahalo tsa tlhahiso ea nucleic acid le sephetho sa tlhoekisos
① Ho netefatsa botšepehi ba sebopeho sa mantlha sa nucleic acid.
② Fokotsa tšitiso ea liprotheine, tsoekere, lipids le li-macromolecule tse ling
③ Ha hoa lokela ho ba le solvent ea tlhaho kapa li-ion tse ngata tsa tšepe tse ka thibelang enzyme ho lisampole tsa nucleic acid.
④ RNA le litšila tse ling tsa nucleic acid li lokela ho felisoa ha ho ntšoa DNA, le ka tsela e fapaneng.
Nako ea poso: Nov-24-2022
