RNAprep Tissue Pusa
Features
■ Faʻalauteleina faʻamaʻa mo meaola o manu e faʻafaigofie ma faigofie ai le faʻagasologa.
■ Tulaga Faʻapitoa DNase Ou te faʻaitiitia genomic DNA faʻaleagaina.
■ O le maualuga-mama ma sauni-e-faʻaaoga RNA e talafeagai mo maaleale i lalo ifo apalai.
■ Leai se phenol / chloroform e toʻesea mai, leai se LiCl ma le ethanol faʻapipiʻiina, ma leai se CsCl gradients centrifugation e manaʻomia, lea e mafai ai ona saogalemu ma faʻatuatuaina le gaioiga.
Talosaga
■ RT-PCR.
■ Itu i Matu, Dot Blot.
■ Taimi Moni PCR.
■ Suʻesuʻega o Chip.
■ PolyA Screening, in vitro faaliliuga, RNase puipuiga faʻapitoa ma molikalone faʻapipiʻi.
Uma oloa mafai ona faʻapitoa mo ODM / OEM. Mo auiliiliga,faʻamolemole kiliki Customized Service (ODM / OEM)
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Mea: 20 mg embryo (13 aso), 15 mg fatugaʻo, 10 mg ate, 15 mg spleen, 10 mg thymus, 20 mg māmā Metotia: Aofaʻiga o le RNA mai ituaiga eseese o isumu na faʻamamaina e faʻaaoga ai le RNAprep Pure Tissue Kit. Iʻuga: O le agarose gel electrophoresis ata o loʻo faʻaalia i luga. 2-4 μl o le 100 μl eluates na utaina ile laina. M: TIANGEN DNA Faʻailoga III; Ala 1-2: Embryo (13 aso); Ala 3: Fatugao; Ala 4-6: ate; Alalaupapa 7: Spleen; Ala 8: Lau Afioga; Ala 9: Lung. O le electrophoresis na faia i le 6 V / cm mo le 30 min i le 1% agarose gel .. |
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).
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