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Ku saabsan soo saarista RNA

Soddon sano ka hor, soo saarista RNA, waxa aan u malayn karno waxay ahayd kaliya sida loo helo RNA dhamaystiran, si kastaba ha ahaatee, ma jirin wax shuruud ah oo loogu talagalay xawaaraha soo saarista (Tiknoolajiyada jiilka kowaad ee habka Trizol ee dhashay).Cusboonaysiinta tignoolajiyada soo saarista RNA, waxa aan samayn karno kuma koobna helitaanka natiijooyinka.Marka ay soo wajahdo soo saarista jiilka kowaad ee reagents-ku-salaysan ee Trizol, tijaabiyeyaashu guud ahaan way dhibeen talaabooyinkeeda qallafsan;in kastoo wanaajin iyo halabuuro badan ka dib, haddana waxay qaadatay waqti badan.Isla mar ahaantaana, reagents organic sida phenol iyo chloroform waxay u baahan yihiin in loo isticmaalo habka soo saarista, taas oo sidoo kale khatar gelinaysa badbaadada shaqaalaha ay khusayso ilaa xad.

2Wax ku ool ah, habboon oo ammaan ah

Helitaanka sheyga soo saarista RNA wax ku ool ah, habboon oo badbaado leh ayaa noqotay yoolka koowaad ee tijaabiyaasha.Sidaa darteed, habka xayeysiiska Silica Spin Column ee soo saarista RNA ayaa la sameeyay.Foregene jiilka saddexaad ee RNA wax soo saarka ayaa soo jiidanaya faa'iidooyinka jiilkii hore ee alaabta soo saarista tiirarka, iyadoo la adeegsanayo (DNA-Nadiifinta + RNA kaliya) laba tikniyoolajiyada Spin Column si loo saaro gDNA iyo haraaga unugyada isku mar, iyada oo aan la daaweyn DNA-da haraaga gDNA, oo aan la isticmaalin phenol, iwm.

Shaxda isbarbardhigga habka hawlgalka

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Shaxda isbarbardhigga natiijooyinka tijaabada

Qalabka go'doominta RNA ee Foregene wuxuu leeyahay wax soo saar sare oo RNA ah, nadiif wanaagsan, iyo saamaynta soo saarista miRNA ayaa ka fiican xirmooyinka kale ee astaanta.

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Electrophoresis of 50 mg ee beerka jiirka wadarta RNA soosaaray habab kala duwan
Jiilka koowaad (1: Trizol)
Jiilka labaad (2: Qalabka soo saarista RNA ee shirkadda A)
Jiilka sadexaad

garaafka qPCR (garaafka qalooca kor u qaadista)

DNA-da genomic waa in si wax ku ool ah looga saaraa inta lagu jiro soo saarista RNA, haddii kale waxay yeelan doontaa saameyn aan la xakamayn karin oo ku saabsan tijaabooyinka hoose (qiimaha hore ee CT, faa'iidooyinka beenta ah).Xirmada jiilka soo socda ee Foregene ayaa si fiican u saari kara DNA-da genomic iyada oo aan lagu darin DNA-da, badbaadinta waqti badan, iyada oo laga fogaanayo hoos u dhaca ay keento wasakheynta RNase-ka baxsan.
Jiilka koowaad (cagaarku wuxuu tilmaamayaa TRlzol-, casaanku wuxuu muujinayaa TRlzol+)

5Jiilka labaad (Buluugu waxa uu tilmaamayaa Shirkadda A xirmada soo saarista RNA -, casaanku waxa uu tilmaamayaa shirkadda A RNA xirmada +)

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Jiilka saddexaad (buluug ayaa tilmaamaya shirkadda A RNA xirmada soo saarista oo lagu daray, cagaarka ayaa muujinaya xirmooyinka Go'doominta RNA ee Foregene)

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(Fiiro gaar ah: "_" garaafka qPCR wuxuu muujinayaa in DNA-da aan lagu darin, iyo wasakheynta DNA-da aan la saarin; "+" waxay muujineysaa in DNA lagu daray, iyo wasakheynta DNA-da laga saaray)

Ku dhammaystir soo saarista RNA 11 daqiiqo gudahood

Looma baahna sanado badan oo ururinta khibrada soo saarista aashitada nucleic-ka, Foregene wax-ku-oolnimada sare ee wadarta guud ee xirmooyinka go'doominta RNA waxay kuu oggolaan kartaa inaad ku hesho RNA-soo-saar sare iyo saafi-sare 11 daqiiqo gudahood si aad ula kulanto baahiyo kala duwan oo tijaabo ah.

9Ilaa hadda, Foregene wadarta guud ee xirmooyinka isolaton ee taxanaha alaabada waxaa aqoonsaday tiro badan oo macaamiil ah waxayna gacan ka geysteen suugaan badan oo dhibcaha sare leh.Isticmaalka dhabta ah ee macaamilka ee jaantuska elektiroonigga ah ee jawaab celinta iyo qalooca korantada PCR ee tirada ka dib qoraal-ku-soo-noqoshada ayaa muujinaya in RNA laga soo saaray isticmaalka alaabta Foregene kaliya maaha mid aad u sarreeya, laakiin sidoo kale waa daacadnimo wanaagsan.Kaliya tignoolajiyada wax soo saarka ee lagu tijaabiyay suuqa ayaa qof walba u roonaan karaa.

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Qalabka Go'doominta unugga ee RNA

Wax soo saarka RNA sare

Dhakhso: Ku dhame go'doominta RNA 11 daqiiqo gudahood

Badbaadada: Wax kiimiko ah oo dabiici ah laguma darin

Qaar ka mid ah xigashooyinka dhibcaha sare leh:

1. Yuan Fang, Zezhong Liu, Yang Qiu, iyo al.Joojinta fayras-celinta borotiinka RNAi ee naqshadeeyaha peptides wuxuu ka ilaaliyaa caabuqa enteroviral ee vivo, Difaaca, 54 (10), 2021: 2231-2244.e6.(IF:31.745,Qalabka Go'doonka ah ee RNABisad No.RE-02011)

https://www.sciencedirect.com/science/article/pii/S1074761321003617

2. Ren, Y., Wang, A., Wu, D. iyo al.Laba-ka-hortagga difaaca gudaha iyo apoptosis ee borotiinka cytomegalovirus ee bini'aadamka UL37x1 waxay awood u siinaysaa fayraska ku-noqoshada hufan.Nat Microbiol 7, 1041-1053 (2022).(IF:30.964,Qalabka Go'doominta unugga ee RNABisad No.RE-03111)

https://doi.org/10.1038/s41564-022-01136-6


Waqtiga boostada: Jul-27-2022