Bipụtara: CAS Key Laboratory of Pathogen Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences
Ọ bụrụ: 69.504
Ngwaahịa enyere aka:Ngwakọta mkpokọta cell RNA iche
Njikọ izizi:https://www.nature.com/articles/s41586-022-05271-2
Ọrịa na filoviruses dị ka Ebola virus (EBOV) na Marburg nje nwere ike ịkpata nnukwu mgbaàmà ụlọ ọgwụ, gụnyere hemorrhagic ahụ ọkụ na multiorgan ọdịda.4.N'ebe a, anyị kpebisiri ike nhazi nke nje oria Ebola L na mgbagwoju anya na tetrameric VP35 site na iji microscopy cryo-electron (steeti 1).Nnyocha nhazi gosipụtara na nje Ebola L nwere ihe ntinye ntinye nke filovirus nke dị mkpa maka njikọ RNA, yana na VP35 na-emekọrịta ihe na mpaghara N-terminal L site na atọ protomers nke VP35 tetramer.
N'ime akwụkwọ a, ejiri Foregene wepụta RNA niileNgwakọta mkpokọta cell RNA ichena Nanodrop tụrụ ntinye nke RNA na 260 nm, wee mee nchọpụta RT/qPCR.
■ A na-arụ ọrụ ahụ dum na ụlọ okpomọkụ (15-25 ℃), na-enweghị ice bat na obere okpomọkụ centrifugation.
■ Ngwa niile enweghị RNase, ọ nweghị mkpa ichegbu onwe gị maka mmebi RNA.
∎ kọlụm DNA- Cleaning na-ejikọta DNA kpọmkwem, ka ngwa ahụ wee nwee ike wepụ nje DNA nje na-etinyeghị DNA ọzọ.
∎ Mkpụrụ RNA dị elu: Kọlụm naanị RNA nwere ike sachaa RNA nke ọma.
∎ Ọsọ ọsọ: dị mfe iji rụọ ọrụ ma nwee ike mechaa n'ime nkeji iri na otu.
■ Nchekwa: Ọ dịghị ihe reagent organic achọghị.
■ Ọdịmma dị elu: RNA dị ọcha dị oke ọcha, enweghị protein na adịghị ọcha ndị ọzọ, ọ nwekwara ike izute nnwale dị iche iche na-esote.
Oge nzipu: Nov-04-2022
