■ Maualuga fetuʻutuʻunaʻi: vao vao o amata faʻataʻitaʻiga voluma, talafeagai mo tele lapisi faʻataʻitaʻiga toʻesea i se tasi tali atu.
■ Maualuga maualuga: Faʻasologa auala faʻateleina le faʻatupuina o RNA i faʻataʻitaʻiga.
■ Lautele faʻaaoga: E talafeagai mo le tele o faʻataʻitaʻiga eseese e pei o laʻau ma manu, sela tuʻufaʻatasia, toto, suavai o le tino, ma isi.
■ Faagaioiga vave: Genomic DNA mafai ona maua i totonu o le 1 itula ..
Ituaiga: Faʻasologa faʻavae faʻavae
Faataʻitaʻiga: Siama, siama, siama, meaola, laʻau toto, sela faʻaleaganuʻu ma sua o le tino.
Taunuʻuga: RNA
Taimi o galuega: ~ 1 itula
Talosaga: TRNzol Universal reagent faʻaitiitia le faʻaleagaina o faʻaletino e pei o DNA ma polotini i le faʻamamaina o le RNA, ma e mafai ona faʻaaoga saʻo mo faʻataʻitaʻiga biology biology eseese pei o Northern Blot, Dot Blot, PolyA faʻataʻitaʻiga, i vitro faʻaliliuga, RNase puipuiga puipuiga, fausiaina faletusi CDNA , RT-PCR, taimi moni PCR ma le maualuga-throughput faʻasologa.
Uma oloa mafai ona faʻapitoa mo ODM / OEM. Mo auiliiliga,faʻamolemole kiliki Customized Service (ODM / OEM)
Metotia: 30 mg rat ate vaega, 100 mg araisa laʻau na aoina e suavai nitrogen olo; 1 × 106HepG2 siʻosiʻomaga sela ma 700 μl Saccharomyces Cerevisiae aganuu auala (OD600 = 0.9) na aoina e centrifugation. 1 ml o TRNzol Universal Reagent mai TIANGEN ma oloa talafeagai mai le kamupani L ma le T na faʻaopopoina i aliquot taʻitasi o faʻataʻitaʻiga ma le RNA aveʻeseʻese na faia i le mulimuli ai i faʻatonuga na saunia e kamupani taʻitasi. O le eluma voluma sa 80 μl, 50 μl, 30 μl ma 30 μl mo le fa faʻataʻitaʻiga faʻatulagaina. 3 μl o le eluate na utaina ile laina.
MIII: TIANGEN Faailoga III;
O le electrophoresis na faia i le 6 V / cm mo le 30 min i luga o le 1% agarose.
Iʻuga: TIANGEN TRNzol Universal Reagent mafai ona aveʻese maualuga mama ma lelei amio faʻamaoni RNA mai isumu ate, laisa araisa, sela tuʻufaʻatasia ma fefete faʻataʻitaʻiga, ma maualuga lelei. O le lelei RNA e faʻatusatusa pe iʻu teisi maualuga foi nai lo le sapalai L ma T oloa.
A-1 Cell lysis poʻo le homogenization le lava
---- Faʻaititia faʻataʻitaʻiga faʻaaogaina, faʻateleina le aofaʻi o lysis buffer, faʻateleina homogenization ma lysis taimi.
A-2 Sample aofaiga e telē tele
---- Faʻaititia le aofaʻi o faʻataʻitaʻiga faʻaaogaina pe faʻateleina le aofaʻi o lysis buffer.
A-1 Le lava le sela lysis poʻo le homogenization
---- Faʻaititia faʻataʻitaʻiga faʻaaogaina, faʻateleina le aofaʻi o lysis buffer, faʻateleina homogenization ma lysis taimi.
A-2 Sample aofaiga e telē tele
---- Faʻamolemole vaʻai i le maualuga gafatia gafatia.
A-3 RNA e le elied atoa mai le koluma
---- A maeʻa ona faʻaopopo le RNase-Free vai, tuʻu mo ni nai minute ae le i totogia le centrifuging.
A-4 Ethanol i le eluent
---- A maeʻa le mulumulu, toe faʻatulaga lelei ma aveese le faʻamago i le mea sili e mafai.
A-5 Cell aganuu auala e le maeʻa uma
---- Pe a faʻaputuputuina sela, faʻamolemole ia mautinoa ia aveʻese le aganuʻu auala i le tele e mafai ai.
A-6 O sela o loʻo teuina ile RNAstore e leʻo totogia lelei
---- RNAstore density e sili atu nai lo le averesi sela aganuu auala; o lea e tatau ona faʻateleina le centrifugal force. E fautuaina e centrifuge ile 3000x g.
A-7 Low RNA mea ma tele i le faʻataʻitaʻiga
---- Faʻaaoga se faʻataʻitaʻiga lelei e faʻamautinoa ai pe o le maualalo o fua e mafua mai i le faʻataʻitaʻiga.
A-1 O mea e le fou
---- O mea fou e tatau ona teuina i totonu o le vai nitrogen i le taimi lava lena poʻo le vave tuʻuina i totonu o le RNAstore reagent e mautinoa ai le aveʻeseina aʻafiaga.
A-2 Sample aofaiga e telē tele
---- Faʻaititia le aofaʻi aofaʻi.
A-3 RNase faʻaleagainan
---- E ui lava e leʻo iai le RNase i le buffer o loʻo i totonu o le pusa, e faigofie lava ona faʻaleagaina le RNase i le taimi e aveese ai ma e tatau ona faʻatautaia ma le faʻaeteete.
A-4 Faʻaleagaina eletise
---- Sui le paipa eletise ma ia mautinoa o mea faʻaaoga ma Uta Faʻafefe e leai se faʻamaʻi RNase.
A-5 Tele tele utaina mo electrophoresis
---- Faʻaitiitia le aofaʻi o faʻataʻitaʻiga utaina, o le utaina o vaiʻeli uma e le tatau ona sili atu i le 2 μg.
A-1 Faʻataʻitaʻiga aofaʻi e telē tele
---- Faʻaititia le aofaʻi aofaʻi.
A-2 Nisi faʻataʻitaʻiga e maualuga aano o DNA ma e mafai ona togafitia ile DNase.
---- Faia togafitiga RNase-Free DNase ile tali RNA na maua, ma e mafai ona faʻaaoga saʻo le RNA mo suʻesuʻega mulimuli ane pe a maeʻa togafitiga, pe mafai foi ona faʻamamaina e RNA faʻamamaina ato.
Mo tioata, tao i le 150 ° C mo le 4 h. Mo palasitika koneteina, faʻatofu i le 0.5 M NaOH mo le 10 min, ona fufulu faʻamaeʻa lea i le RNase-leai se vai ona faʻamamaina lea e aveʻese atoa le RNase. O le reagents poʻo vaifofo faʻaogaina i le faʻataʻitaʻiga, aemaise lava le vai, tatau ona saoloto mai RNase. Faʻaaoga le vai e leai se RNase mo sauniuniga uma (faʻaopopo vai i se fagu tioata mama, faʻaopopo le DEPC i le faʻaiʻuga mulimuli o le 0.1% (V / V), lulu le po ma le autoclave).
Talu mai lona faavaeina, o lo tatou fale gaosi ua atiina ae muamua lalolagi vasega oloa ma mulimulitaia le mataupu faavae
o lelei muamua. Tatou oloa ua maua sili lelei taʻuleleia i le alamanuia ma valuabletrusty i fou ma tuai tagata faʻatau ..