Vortexing yuav tsis txiav koj RNA ua rau nws siv tsis tau yog tias koj tab tom ua random priming RT - tsuas yog koj xav clone ntev mRNA. Kev txhawb nqa Invitrogen tech yog qhov tseeb thiab. Chloroform hnyav dua Trizol thiab yuav tog rau hauv qab ntawm lub raj.
Nws puas nyab xeeb rau vortex RNA?
Tsis txhob vortex Trizol lysates lossis RNA cov qauv kom tsis txhob shearing. Tom qab rho tawm, khaws cov qauv RNA ntawm cov dej khov txhua lub sijhawm. Cov txheej txheem no yog tsim los rau cov qauv lysed 1mL ntawm Trizol hauv 1.5 lossis 2mL raj.
Koj puas tuaj yeem sov RNA?
Yog li ntawd, tsis txhob kub siab ( saum toj +65 ° C) vim cov no cuam tshuam rau kev ncaj ncees ntawm RNA. Hloov chaw, kom yaj tawm cov txheej txheem thib ob, cua sov RNA rau +65 ° C rau 15 feeb nyob rau hauv lub xub ntiag ntawm denaturing buffers.
Yuav ua li cas koj resuspend RNA pellets?
resuspend koj cov pellet nrog dej sov (40-50 deg. celc). incubate hauv 45 deg celc rau 10 mins. muab lawv in +4 rau 4-6 teev.
Koj ua nag cua RNA li cas?
A.
Tom qab cov ntsev concentration tau hloov kho, RNA yuav raug los ntawm ntxiv 2.5 ntim ntawm ethanol lossis 1 ntim ntawm isopropanol thiab sib tov kom zoo, ua raws li txias txias li 15 feeb ntawm -20 ° C.
