Kungakanani okwaziyo

Mayelana no-Nukukhishwa kwe-ucleic acid

Ukuqala nokuphela kwe-nucleic acid ehlanzekile

Konke kunzima ekuqaleni, i-nucleic acid ehlanzekile iyona kakhulu

Ukuqaliswa kokuhlolwa kwamangqamuzana, kokuhlolwa okulandelayo

Impumelelo noma ukwehluleka kunomthelela obalulekile.

I-spectrophotometer ye-Trace/ultra-trace ye-UV ithandwa abacwaningi abaningi besayensi ngoba ikwazi ukubona ukugxiliswa kwe-nucleic acid kanye nezinsalela ze-ion ye-usawoti kalula, ngokushesha nangokongayo.

Njengoba sazi sonke, i-A260 isho inani le-OD lokumunca i-nucleic acid, i-A280 isho inani le-OD lokumuncwa kwe-protein, i-A230 isho inani le-OD lokumunca i-ion kasawoti, ngakho i-A260 ingakwazi ukuguqula ukuhlushwa kwe-nucleic acid, i-A260/280 isho izinsalela zamaphrotheni, i-A260/230 isho i-ion ion kasawoti etholwe, kodwa lokhu kuyinsalela esekelwe ocwaningweni.ezejwayelekile” ukukholelwa ukuthi ingachaza ubumsulwa be-DNA ne-RNAngezinga elithile.Akuyona indinganiso kuphela yokuhlonza isivuno se-nucleic acid nobumsulwa, isetshenziswa kuphela njengereferensi, futhi ayiyona "izinga legolide".

Imanuwali ye-Thermo Fisher ND-2000 igcizelela ukwethembeka kwemiphumela yokuhlolwa

Isizathu siwukuthi imiphumela etholwe kusetshenziswa i-micro/ultramicro UV spectrophotometer ibonisa kuphela isivuno nobumsulwa be-nucleic acid ohlangothini, futhi kunezici eziningi ezinomthelela (indlela yokubona, isampula yevolumu, ukugxiliswa kwesampula, inani le-pH, amanye ama-ion athinta ukulinganiswa kokumunca, njll.) , inani le-OD elilinganisiwe alinembile ngempela.Ngesikhathi esifanayo, ngenxa yokuntuleka kokucaciswa kokunqunywa kwenani lokumunca, i-DNA enomucu owodwa, i-DNA enemicu ephindwe kabili, i-RNA, ama-dNTP, namanye amaprotheni anezinga eliphakeme lokumunca ku-260nm wavelength, futhi ukugxila okunqunywa i-A260 iyodwa akusho ukuthi i-DNA yangempela noma i-RNA.Ukugxilisa ingqondo, kanye nobuqotho nokwehliswa kwayo angeke kwahlulelwe.

Ngakho-ke singahlulela kanjani ikhwalithi ye-nucleic acid yethu ehlanzekile?Ngaphezu kokusebenzisa i-spectrophotometer ye-UV encane/ultramicro ukuze kutholwe, izindlela ezifana ne-agarose gel electrophoresis nazo ziyadingeka ukuze zibonise ubumsulwa nobuqotho be-nucleic acid, futhi zibonise ukugxilisa ingqondo ngezinga elithile.Ngale ndlela, isivuno se-nucleic acid nokuhlanzeka okutholwe emiphumeleni yokutholwa yezindlela ezahlukahlukene kukholakala.

Ukuqhathaniswa kweshadi lokuhlola

I-Genomic DNA yamaseli e-H1299 yakhishwa kusetshenziswa amakhithi enkampani A's DNA Extraction, futhi kwabonakala ukungcoliswa kokungcola okuphawulekayo, okubangele amanani aphezulu e-OD.

(Inani lokulinganisa i-OD kanye ne-electropherogram ehambisanayo)

Inkomba yokuhlola

Ingabe likhona “izinga legolide” lesivuno se-nucleic acid nokuhlolwa kwekhwalithi?

Ngokwazi kwethu, ayikho indlela elula, esheshayo futhi eshibhile.Kungakhathaliseki ukuthi i-spectrophotometer, i-gel electrophoresis, noma i-mass spectrometry, zonke zibonisa ukugxila nokuhlanzeka kwe-nucleic acid esixazululweni esivela ezicini ezihlukahlukene, futhi zidinga ukwahlulelwa ngokubhekisela komunye nomunye.

Inkomba yokuhlola engcono kakhulu ihlala njalouhlelo lokusebenza lokuhlola lokulandelela.Akuthinti ukusebenza kahle kokuhlehla kokuloba kanye nokukhulisa i-PCR.Ukuthola imikhiqizo yokukhulisa i-amplification enokwethenjelwa kanye namanani e-Ct, kanye nokuthola ukulandelana okuphelele ngokulandelanisa kuwumphumela wokukhipha i-nucleic acid eyimpumelelo.

Ukufingqa, umbono wetiyori yesilinganiso kuphela kufanele kuphikiswe umbono "wokusebenzisa imiphumela emihle yokuhlola eya phansi njengamapharamitha amahle okukhipha"!

Okunconyiwe kwekhithi yokukhipha i-Foregene DNA RNA ukuze uthole ukuhlanzeka okuphezulu kwe-DNA/RNA:

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

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