Yeyiphi indlela yokwahlula nokucoca ii-nucleic acids?

Ii-asidi ze-nucleic, kuquka i-DNA kunye ne-RNA, zii-biomolecule ezibalulekileyo ezidlala indima ebalulekileyo kwi-genetics, i-molecular biology, kunye ne-biotechnology. Ukukwazi ukwahlula nokucoca ezi nucleic acids kubalulekile kwiintlobo ngeentlobo zezicelo, kuquka ukuqokelela, ukulandelelana, kunye nohlalutyo lokubonakaliswa kwezakhi zofuzo. Iinkqubo zokucoca i-Nucleic acid ziquka uluhlu lweendlela ezenzelwe ukukhupha nokucoca ii-nucleic acids kwiisampuli zebhayoloji. Eli nqaku lihlola iindlela zokwahlula nokucoca ii-nucleic acids kwaye ligxininisa ukubaluleka kwezi nkqubo kuphando lwesayensi lwanamhlanje.

Ukuqonda ukucocwa kwe-nucleic acid

Ukuhlanjululwa kwe-Nucleic acid kubhekisa ekukhutshweni kwe-DNA okanye i-RNA kwiiseli okanye kwizicubu, kulandele ukususwa kwezinto ezingcolisayo ezifana neeproteni, ii-lipids, kunye nezinye izinto ezingafunekiyo kwiiseli. Ubunyulu kunye nokuthembeka kwee-nucleic acids ezizimeleyo kubalulekile kwizicelo ezisezantsi, njengoko ukungcola kunokuthintela iimpendulo ze-enzyme kwaye kuchaphazele ukuchaneka kweziphumo zovavanyo.

Iindlela eziqhelekileyo zokwahlulwa kunye nokucoca i-nucleic acid

Ukukhutshwa kwePhenol-chloroform:Le ndlela yendabuko isebenzisa izinyibilikisi ze-organic ukwahlula ii-nucleic acids kwiiproteni kunye nezinye izinto zeseli. Isampuli ixutywe ne-phenol kunye ne-chloroform, nto leyo ebangela ukuba ii-nucleic acids zahlulwe zibe yi-aqueous phase, ngelixa iiproteni zihlala zikwi-organic phase. Emva kwe-centrifugation, i-aqueous phase equlethe ii-nucleic acids iyaqokelelwa kwaye ixutywe ne-ethanol.

Iindlela ezisekelwe kwijeli yeSilica:Iimembrane zejeli ye-silica zisetyenziswa kakhulu kwiikiti zokucoca i-nucleic acid zorhwebo. Umgaqo wale ndlela kukuba ii-nucleic acids zibophelele kwi-silica gel xa zinomlinganiselo ophezulu wetyuwa. Emva kokubopha, izinto ezingcolisayo ziyahlanjwa, emva koko ii-nucleic acids ziyasuswa nge-buffer enetyuwa encinci okanye amanzi. Le ndlela iyathandwa kuba ikhawuleza, isebenza kakuhle, kwaye ivelisa ii-nucleic acids ezicocekileyo kakhulu.

Ukucocwa kweentsimbi zeMagnetic:Le ndlela isebenzisa iintsimbi zemagnethi ezigqunywe ngee-nucleic acid binding agents. Xa isampuli ixutywe neentsimbi zemagnethi, ii-nucleic acids zifunxa phezu kweentsimbi. Iintsimbi zahlulwe kwisisombululo kusetyenziswa imagnethi, ngaloo ndlela zisusa ungcoliseko. Le ndlela inokusetyenziswa ngeendlela ezahlukeneyo kwaye inokwenziwa ngokuzenzekelayo, nto leyo eyenza ukuba ifaneleke kwizicelo eziphezulu.

I-chromatography yeekholamu:Le ndlela ibandakanya ukudlulisa isampuli kwikholamu ye-chromatographic egcwele isigaba esingaguqukiyo esigcina ii-nucleic acids ngokukhetha. Iindidi ezahlukeneyo zeekholamu zingasetyenziswa, kuquka ezo zisekelwe ekukhutshweni kobungakanani okanye kwimigaqo yokutshintshiselana kwee-ion. Ii-nucleic acids ziyaphuma kwikholamu, nto leyo ephumela kwisampuli ecocekileyo.

Iindlela ze-enzyme:Iindlela ze-enzyme, ezifana nezo zisebenzisa i-DNase okanye i-RNase, zingasetyenziselwa ukutshabalalisa ii-nucleic acids okanye izinto ezingcolisayo ezingafunekiyo. Le ndlela isebenza ngakumbi xa kusetyenzwa iisampulu ezintsonkothileyo, ezifana nezo ziqulethe i-DNA kunye ne-RNA.

Ukuququmbela

Ukwahlulwa kunye nokucocwa kwe-nucleic acid ngamanyathelo abalulekileyo kuphando lwebhayoloji yeemolekyuli kunye nokusetyenziswa kwayo.Iinkqubo zokucoca i-nucleic acidbanika abaphandi iindlela ezahlukeneyo zokufumana ii-nucleic acids ezikumgangatho ophezulu ezifanelekileyo kwizicelo ezisezantsi. Nokuba kusetyenziswa i-phenol-chloroform extraction yendabuko okanye iindlela zanamhlanje ezifana ne-silica gel okanye i-magnetic bead cleansing, ukhetho lwendlela luxhomekeke kwiimfuno ezithile zovavanyo kunye nohlobo lwesampulu. Ngokuhambela phambili kwetekhnoloji, ezi nkqubo zokucoca ziye zaguquka ngokuqhubekayo, ziphucula ukusebenza kakuhle, isantya, kunye nokuthembeka, ekugqibeleni ziphucula amandla abaphandi kwicandelo lebhayoloji yemolekyuli.