50 Imyiteguro, 200 Imyiteguro

Iki gikoresho gikoreshakuzenguruka inkingi na formulayatejwe imbere nisosiyete yacu, ishobora gukuramo isuku nini kandi yujuje ubuziranenge RNA yose mubice bitandukanye byinyamanswa kandi ikora neza.Bitanga inkingi ikora neza ya ADN-isukura, ishobora gutandukanya byoroshye kandi ikanatanga ADN ya genomic na lysate ndengakamere na tissue, byoroshye no guta igihe;Inkingi ya RNA yonyine irashobora guhuza RNA neza kandi irashobora gutunganyirizwa icyarimwe hamwe na formula idasanzwe Ingero nyinshi.

Sisitemu yose ni RNase-Yubusa, kugirango RNA yakuweho itangirika;Buffer RW1, Buffer RW2 sisitemu yo gukaraba, kugirango RNA yabonetse idafite proteyine, ADN, ion, hamwe n’umwanda wanduye.

Ibigize ibikoresho

Amakuru y'ibicuruzwa

Ibiranga ibyiza

■ Ntibikenewe ko uhangayikishwa no kwangirika kwa RNA;sisitemu yose ni RNase-Ubuntu
Kuraho neza ADN ukoresheje ADN-Isukura Inkingi
Kuraho ADN utongeyeho DNase
Ibikorwa byoroshye-byose birangirira ku bushyuhe bwicyumba
■ Kwihuta -ibikorwa birashobora kurangira muminota 30
■ Umutekano-nta reagent ukenewe
Pure Isuku ryinshi -OD260 / 280≈1.8-2.1

Porogaramu

Birakwiriye gukuramo no kweza RNA yose mubice bitandukanye byinyamanswa nshya cyangwa bikonje cyangwa ingirabuzimafatizo.

Ibipimo byibicuruzwa

Applices Porogaramu yo hasi: synthesis ya mbere ya cDNA, RT-PCR, clone ya molekile, Blot y'Amajyaruguru, nibindi.
Ingero: inyama zinyamanswa, ingirabuzimafatizo
Ingano: Tissue 10-20mg, Ingirabuzimafatizo (2-5) × 10⁶
Ubushobozi ntarengwa bwa ADN bwo guhuza inkingi yo kweza: 80 μg
Volume Ingano yo gukuraho: 50-200 μl

Urujya n'uruza rw'akazi

Amatungo Yuzuye RNA Yigunze Igikoresho cyavuwe 20mg
Icyitegererezo cyimbeba, fata 5% isukuye yose RNA 1% agar

Glycogel electrophoreis
1: Impyiko 2: Impyiko
3: Umwijima 4: Umutima

Ububiko nubuzima bwiza

Igikoresho gishobora kubikwa amezi 24 mubushyuhe bwicyumba (15-25 ℃) cyangwa 2-8 ℃ mugihe kirekire.Buffer RL1 irashobora kubikwa kuri 4 ℃ ukwezi 1 nyuma yo kongeramo merc mercaptoethanol (bidashoboka).

 

Ingingo zavuzwe

1.NIBA: 18.808:Zheng, Q., Qin, F., Luo, R., n'abandi.mRNA-Yuzuye Lipid-Nka Nanoparticles yo Guhindura Umwijima Binyuze muri Optimisation yo Hagati Igishushanyo mbonera.Umujyanama.Imikorere.Ibikoresho.2021, 31, 2011068.doi: 10.1002 / adfm.202011068.

2.NIBA: 18.187:We X, Hong W, Yang J, n'abandi.Apoptose ya selile mu buryo bwo kuvura ingirangingo ngengabuzima itegura igira ingaruka zo gukingira indwara binyuze mu kurekura fosifatiqueylserine.Ikimenyetso cyo Gutwara Intego Intego.2021 Nyakanga 14; 6 (1): 270.doi: 10.1038 / s41392-021-00688-z.

3.NIBA: 17.97 ai Z, Liu H, Liao J, n'abandi.N7-Methylguanosine tRNA guhindura byongera ubusobanuro bwa oncogenic mRNA kandi biteza imbere cholangiocarcinoma intrahepatic.Akagari ka Mol.2021 Nyakanga 29: S1097-2765 (21) 00555-4.doi: 10.1016 / j.molcel.2021.07.003.

4.NIBA: 9.225: Cao X, Shu Y, Chen Y, n'abandi.Mettl14-Yunganirwa na m6A Guhindura byorohereza kuvugurura umwijima ukomeza Endoplasmic Reticulum Homeostasis.Akagari Mol Gastroenterol Hepatol.2021; 12 (2): 633-651.doi: 10.1016 / j.jcmgh.2021.04.001.

 

RNA kwigunga ibikoresho bya izindi ntangarugerozirahari:

Akagari, ibimera, virusi, amaraso, nibindi

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Mbere: Akagari ka Direct RT QPCR Kit-SYBR GREEN I.

Ibikurikira: Uruganda rukora firime irwanya izuba

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