Iyiphi indlela yokuhlukanisa nokuhlanza ama-nucleic acid?

Ama-asidi e-nucleic, okuhlanganisa i-DNA ne-RNA, yi-biomolecule ebalulekile edlala indima ebalulekile ku-genetics, i-molecular biology, kanye ne-biotechnology. Ikhono lokuhlukanisa nokuhlanza lawa ma-nucleic acid libalulekile ezinhlotsheni ezahlukene zokusebenza, okuhlanganisa ukwakheka kwe-cloning, ukulandelana, kanye nokuhlaziywa kokubonakaliswa kwezakhi zofuzo. Izinhlelo zokuhlanza i-nucleic acid zihlanganisa uhla lwezindlela ezenzelwe ukukhipha nokuhlanza ama-nucleic acid kumasampula ebhayoloji. Lesi sihloko sihlola izindlela zokuhlukanisa nokuhlanzwa kwama-nucleic acid futhi siqokomisa ukubaluleka kwalezi zinhlelo ocwaningweni lwesayensi lwanamuhla.

Ukuqonda ukuhlanzwa kwe-nucleic acid

Ukuhlanzwa kwe-Nucleic acid kubhekisela ekukhishweni kwe-DNA noma i-RNA emaseli noma ezicutshini, kulandelwe ukususwa kwezinto ezingcolisayo ezifana namaprotheni, ama-lipid, kanye nezinye izinsalela zamaseli. Ubumsulwa kanye nobuqotho bama-nucleic acid ahlukanisiwe kubalulekile ekusetshenzisweni okulandelayo, njengoba ukungcola kungavimbela ukusabela kwama-enzyme futhi kuthinte ukunemba kwemiphumela yokuhlola.

Izindlela ezivamile zokuhlukanisa nokuhlanza i-nucleic acid

Ukukhishwa kwe-Phenol-chloroform:Le ndlela yendabuko isebenzisa izinyibilikisi eziphilayo ukuhlukanisa ama-nucleic acid kumaprotheni nezinye izingxenye zamaseli. Isampula ixutshwa ne-phenol kanye ne-chloroform, okubangela ukuba ama-nucleic acid ahlukaniswe abe yisigaba samanzi, kuyilapho amaprotheni ehlala esigabeni sezinto eziphilayo. Ngemva kokuxuba, isigaba samanzi esiqukethe ama-nucleic acid siyaqoqwa futhi sifakwe i-ethanol.

Izindlela ezisekelwe kujeli ye-silica:Ama-membrane ejeli le-silica asetshenziswa kabanzi kuma-kit okuhlanza i-nucleic acid kwezohwebo. Isimiso sale ndlela ukuthi ama-nucleic acid abopha ijeli le-silica ngamanani aphezulu kasawoti. Ngemva kokubopha, ukungcola kuyagezwa, bese ama-nucleic acid esuswa nge-buffer noma amanzi anosawoti omncane. Le ndlela iyathandwa ngoba iyashesha, isebenza kahle, futhi ikhiqiza ama-nucleic acid ahlanzekile kakhulu.

Ukuhlanzwa kobuhlalu be-magnetic:Le ndlela isebenzisa ubuhlalu obunobuthi obumbozwe ngezinto zokubopha i-nucleic acid. Uma isampula ixutshwa nobuhlalu obunobuthi, ama-nucleic acid anamathela ebusweni bobuhlalu. Ubuhlalu bube sebuhlukaniswa nesixazululo kusetshenziswa i-magnet, ngaleyo ndlela kususwe ukungcola. Le ndlela isebenziseka ngezindlela eziningi futhi ingenziwa ngokuzenzakalela, okwenza ifaneleke ezinhlelweni zokusebenzisa amandla aphezulu.

I-chromatography yamakholomu:Le ndlela ihilela ukudlulisa isampula kukholomu ye-chromatographic egcwele isigaba esingaguquki esigcina ama-nucleic acid ngokukhetha. Izinhlobo ezahlukene zamakholomu zingasetshenziswa, kufaka phakathi lezo ezisekelwe ekukhishweni kosayizi noma ezimisweni zokushintshana kwama-ion. Ama-nucleic acid ayaphuma kukholomu, okuholela kusampula ehlanziwe.

Izindlela ze-enzyme:Izindlela ze-enzyme, njengalezo ezisebenzisa i-DNase noma i-RNase, zingasetshenziswa ukwehlisa ngokukhetha ama-nucleic acid noma izinto ezingcolisayo ezingafuneki. Le ndlela isebenza kahle kakhulu lapho kucutshungulwa amasampula ayinkimbinkimbi, njengalawo aqukethe i-DNA ne-RNA.

Ekuphetheni

Ukuhlukaniswa kanye nokuhlanzwa kwe-nucleic acid kuyizinyathelo ezibalulekile ocwaningweni lwebhayoloji yama-molecule kanye nokusetshenziswa kwawo.Izinhlelo zokuhlanza i-nucleic acidnikeza abacwaningi izindlela ezahlukahlukene zokuthola ama-nucleic acid asezingeni eliphezulu afanele ukusetshenziswa ngezansi. Kungakhathaliseki ukuthi kusetshenziswa ukukhishwa kwe-phenol-chloroform yendabuko noma izindlela zesimanje njenge-silica gel noma ukuhlanza okusekelwe ku-magnetic bead, ukukhetha indlela kuncike ezidingweni ezithile zokuhlolwa kanye nohlobo lwesampula. Ngokuthuthuka kwezobuchwepheshe, lezi zinhlelo zokuhlanza ziye zathuthuka ngokuqhubekayo, zithuthukisa ukusebenza kahle, isivinini, kanye nokuthembeka, ekugcineni zathuthukisa amakhono abacwaningi emkhakheni we-molecular biology.