Ni bangahe uzi

Ibyerekeye N.gukuramo aside ucleic

Intangiriro nimpera ya acide nucleic isukuye

Ibintu byose biragoye mugitangiriro, aside nucleic isukuye niyo myinshi

Gutangiza ubushakashatsi bwa molekile, kubushakashatsi bwakurikiyeho

Intsinzi cyangwa gutsindwa bifite ingaruka zikomeye.

Trace / ultra-trace UV spectrophotometer itoneshwa nabashakashatsi benshi mubumenyi kuko irashobora kumenya aside aside nucleique hamwe na proteine ​​hamwe nibisigazwa byumunyu ion byoroshye, byihuse kandi mubukungu.

Nkuko twese tubizi, A260 bivuga aside nucleic acide ya OD agaciro, A280 bisobanura intungamubiri za poroteyine zo mu bwoko bwa OD agaciro, A230 bivuze ko umunyu wa ion ukwirakwiza umunyu wa OD, bityo A260 irashobora guhindura aside nucleic yibanze, A260 / 280 bisobanura ibisigisigi bya poroteyine, A260 / 230 bisobanura ibisigazwa by’umunyu, ariko iri ni itegeko ryavumbuwe n’abashakashatsi bashingiye ku mubare munini wo gupima.bisanzwe”Kwizera ko bishobora gusobanura ubuziranenge bwa ADN na RNA kuriurugero runaka.Ntabwo arirwo rwego rwonyine rwo kumenya umusaruro wa acide nucleique nubuziranenge, ikoreshwa gusa nkurutonde, kandi ntabwo "urwego rwa zahabu".

Igitabo cya Thermo Fisher ND-2000 gishimangira kwizerwa ryibisubizo

Impamvu nuko ibisubizo byabonetse ukoresheje micro / ultramicro UV spectrophotometer yerekana gusa umusaruro nubuziranenge bwa acide nucleic biva kuruhande, kandi hariho ibintu byinshi bigira ingaruka (inzira ya optique, urugero rwicyitegererezo, urugero rwibanze, pH agaciro, izindi ion zigira ingaruka kubipimo byo gukuramo, nibindi), agaciro ka OD gapimwe ntabwo byanze bikunze ari ukuri.Muri icyo gihe, kubera kutagira umwihariko wibikoresho byo kugena agaciro kinjira, ADN imwe imwe, ADN ifite imirongo ibiri, RNA, dNTPs, hamwe na poroteyine zimwe na zimwe zifite impinga zo kwinjirira ku burebure bwa 260nm, kandi kwibanda kuri A260 byonyine ntabwo byanze bikunze ADN cyangwa RNA nyayo.Kwibanda, hamwe nubunyangamugayo no gutesha agaciro ntibishobora gucirwa urubanza.

Nigute dushobora gusuzuma ubwiza bwa acide nucleic yatunganijwe?Usibye gukoresha micro / ultramicro UV spectrophotometer kugirango tumenye, uburyo nka agarose gel electrophoreis burasabwa kandi kwerekana neza ubuziranenge nubusugire bwa acide nucleique, no kwerekana ubunini bwurwego runaka.Muri ubu buryo, aside nucleic itanga umusaruro nubuziranenge bwakuwe mubisubizo byuburyo butandukanye bwizewe.

Kugereranya imbonerahamwe

ADN ya genomic ya selile H1299 yakuwe hifashishijwe ibikoresho bya ADN yo gukuramo ADN ya sosiyete A, kandi hagaragaye umwanda ukomeye wanduye, bivamo indangagaciro nyinshi za OD

(Agaciro ko gupima OD hamwe na electropherogramme ihuye)

Igipimo cyo gusuzuma

Hariho "zahabu" yo gutanga aside nucleic no gupima ubuziranenge?

Nkuko tubizi, nta buryo bworoshye, bwihuse kandi buhendutse.Yaba spekitifotometometero, gel electrophorei, cyangwa mass spectrometrie, byose byerekana kwibanda hamwe nubuziranenge bwa acide nucleique mubisubizo bivuye mubice bitandukanye, kandi bigomba gucirwa urubanza bivugana.

Indangagaciro nziza yo gusuzuma ni buri gihegukurikirana-igeragezwa rya porogaramu.Ntabwo bigira ingaruka kumikorere yo kwandukura no kwongera PCR.Kubona ibicuruzwa byizewe byongerewe agaciro hamwe na Ct indangagaciro, no kubona urutonde rwuzuye mukurikirana ni ugukuramo aside nucleic.

Mu ncamake, igitekerezo cyibipimo-byonyine bigomba gukemurwa no kureba "gukoresha ibisubizo byiza byo hasi yubushakashatsi nkibipimo byiza byo gukuramo"!

Basabwe Foregene ADN ibikoresho byo gukuramo RNA kugirango ubone ADN / RNA isukuye:

https://www.foreivd.com/reagent/dna-isolation-series/

https://www.foreivd.com/reagent/rna-isolation-series/