Wazi kangakanani

Malunga noNukutsalwa kwe-ucleic acid

Ukuqala kunye nokuphela kwe-nucleic acid ecocekileyo

Yonke into inzima ekuqaleni, i-nucleic acid ecocekileyo iyona nto ininzi

Ukuqaliswa kovavanyo lweemolekyuli, kwiimvavanyo ezilandelayo

Impumelelo okanye ukungaphumeleli kunempembelelo ebalulekileyo.

Umkhondo / umkhondo we-UV spectrophotometer ithandwa ngabaphandi abaninzi bezenzululwazi kuba inokubona ukuxinana kwe-nucleic acid kunye neprotheyini kunye neentsalela ze-ion yetyuwa ngokulula, ngokukhawuleza nangoqoqosho.

Njengoko sonke sisazi, i-A260 ithetha i-nucleic acid yokufunxa ixabiso le-OD, i-A280 ithetha i-protein yoxinaniso lwexabiso le-OD, i-A230 ithetha ukuba i-ionconcentration ye-salt absorbance OD value, ngoko ke i-A260 inokuguqula i-nucleic acid concentration, i-A260/280 ithetha intsalela yeprotheyini, i-A260/230 ithetha i-ion yetyuwa, intsalela yetyuwa efunyenweyo, kodwa le mveliso isekelwe kwiziphumo zophando.eqhelekileyo” ukukholelwa ukuba inokucacisa ubunyulu beDNA neRNAumlinganiselo othile.Akuyena kuphela umgangatho wokuchongwa kwesivuno se-nucleic acid kunye nokucoceka, isetyenziswe kuphela njengesalathiso, kwaye akusiyo "umgangatho wegolide".

Incwadi ye-Thermo Fisher ND-2000 igxininisa ukuthembeka kweziphumo zovavanyo

Isizathu kukuba iziphumo ezifunyenwe ngokusebenzisa i-micro/ultramicro UV spectrophotometer ibonisa kuphela isivuno kunye nokucoceka kwe-nucleic acid ukusuka kwicala, kwaye kukho izinto ezininzi ezichaphazelayo (umzila we-optical, umthamo wesampuli, isampuli yoxinaniso, ixabiso le-pH, ezinye iiyoni ezichaphazela ukulinganisa ukufunxa, njl.)Kwangaxeshanye, ngenxa yokunqongophala kokucaciswa kwezinto zokumisela ixabiso lokufunxa, i-DNA enemisonto enye, i-DNA ephindwe kabini, i-RNA, i-dNTPs, kunye nezinye iiprotheni zonke zineencopho zokufunxa kwi-260nm wavelength, kunye nokugxininiswa okugqitywe yi-A260 yodwa akuyomfuneko ukuba i-DNA yokwenene okanye i-RNA.Ukugxila, kunye nengqibelelo kunye nokuthotywa kwayo akunakugwetyelwa.

Ke indlela yokugweba umgangatho we-nucleic acid yethu ecocekileyo?Ukongeza ekusebenziseni i-micro/ultramicro UV spectrophotometer yokukhangela, iindlela ezifana ne-agarose gel electrophoresis nazo ziyafuneka ukuba zibonise ngokubonakalayo ukucoceka kunye nokunyaniseka kwe-nucleic acid, kunye nokubonisa ukugxila kwinqanaba elithile.Ngale ndlela, isivuno se-nucleic acid kunye nococeko olufunyenwe kwiziphumo zokufumanisa iindlela ezahlukeneyo zithembekile.

Uthelekiso lwetshathi yovavanyo

I-Genomic DNA yeeseli ze-H1299 zakhutshwa kusetyenziswa iikhithi ze-A's DNA Extraction yenkampani, kwaye kwabonwa ukungcoliseka kokungcola okubalulekileyo, okubangele amaxabiso aphezulu e-OD.

(Ixabiso lokulinganisa i-OD kunye ne-electropherogram ehambelanayo)

Isalathiso soVavanyo

Ngaba kukho "umgangatho wegolide" wokuvunwa kwe-nucleic acid kunye novavanyo lomgangatho?

Njengoko sisazi, akukho ndlela ilula, ikhawulezayo netshiphu.Ingaba i-spectrophotometer, i-gel electrophoresis, okanye i-mass spectrometry, zonke zibonisa ukugxininiswa kunye nokucoceka kwe-nucleic acid kwisisombululo esivela kwiinkalo ezahlukeneyo, kwaye kufuneka zigwetywe ngokubhekisela komnye nomnye.

Esona salathisi sovavanyo sihlala sihleliulandelelwano lovavanyo lwesicelo.Ayichaphazeli ukusebenza kakuhle kwe-reverse transcription kunye nokukhulisa iPCR.Ukufumana iimveliso ze-amplification ezithembekileyo kunye namaxabiso e-Ct, kunye nokufumana ukulandelelana okupheleleyo ngokulandelelana kuyimpumelelo ye-nucleic acid extraction.

Ukushwankathela, umbono wethiyori yomlinganiselo kuphela kufuneka ucelwe umngeni ngumbono “wokusebenzisa iziphumo zovavanyo ezisezantsi njengeparameters zotsalo ezilungileyo”!

Kucetyiswa iikhithi zokutsalwa kwe-Foregene DNA RNA ukufumana ubunyulu be-DNA/RNA:

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

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