Hordhac:

RNA, magaca sayniska ee ribonucleic acid, soo gaabinta waxay ka timid RiboNucleic Acid, waa nooc ka mid ah acid nucleic ah oo la sameeyay iyadoo la isku xirayo ugu yaraan daraasiin ribonucleotides iyada oo loo marayo bonds fosfodiester.

Ujeedada sahaminta RNA waa in la soo saaro wadarta guud ee RNA ee wasakhda la'aanta ah, iyo mabda'a aasaasiga ah ee soo saarista RNA waxaa lagu soo koobi karaa qiyaas ahaan sida soo socota:

1. Khalkhal geli unugyada marka hore

2. Ka dib ka saar wasakhda makromolecular (protein, lipids, DNA, iwm.)

3. Ugu dambayntii, RNA waa la soo degdegay iyadoo laga saarayo cusbada iyo dareerayaasha organic, oo sii sifeynaya RNA

Hore iyo hadda habka soo saarista

Jiilka kowaad ee habka soo saarista RNA waa habka roobka ee TRIzol (reagents loo isticmaalo waxaa ka mid ah guanidine isothiocyanate, phenol, chloroform, iwm) oo ay alifeen awoowe la yiraahdo Piotr Chomczynski iyo ayeeyo Nicoletta Sacchi, sababtoo ah hindisahani wuxuu ogolaanayaa soo saarista RNA si degdeg ah oo sahlan, tan iyo markaas bilaabay kor u kaca RNA.

Ka dib, awoowe wuxuu iibiyay patentkan Invitrogen, Invitrogen-na wuxuu u dalacsiiyay habkan adduunka oo wuxuu noqday hab si ballaaran loo isticmaalo.

Ka dib, Invitrogen waxaa la wareegay Thermo!

Alaabta matale: Invitrogen's TRIzol reagent

Si kastaba ha ahaatee, habka roobabka TRIzol wuxuu u baahan yahay roobab badan iyo soo saarista RNA, iyo reagents-yada la isticmaalay waa sun waxayna qaataan waqti dheer si ay u shaqeeyaan.

Sidaa darteed, habka sifaynta-jiilka labaad ee tiirka-enzyme ayaa yimid, iyadoo la adeegsanayo xuubka silica gel sida walxo xayeysiis ah oo nucleic ah oo gaar ah si kor loogu qaado soo kabashada RNA ee muunada iyadoo laga saarayo wasakh kale.

Habka jiilka labaad wuxuu fududeeyaa hawlgalka iyadoo lagu saleynayo waxqabadka xasilloon, wuxuu yareeyaa khatarta khatarta ah, wuxuuna wanaajiyaa waxtarka soo saarista RNA.

Alaabta matale: QIAGEN RNeasy iyo inta badan alaabta gudaha

Si kastaba ha ahaatee, hababka jiilka kowaad iyo labaad labaduba waxay u baahan yihiin isticmaalka DNA-ga si meesha looga saaro wasakhowga DNA-da, taas oo u keenta xoogaa dhib ah tijaabooyinka hoose ee xiga.Hadhaaga DNAase waxa ay yaraynaysaa hufnaanta qoraalka gadaasha waxana ay yaraynaysaa xadhkaha kowaad ee cDNA, taas oo saamaynaysa saxnaanta natiijooyinka.

Sidaa darteed, habka soo saarista jiilka saddexaad ayaa la wanaajiyey iyadoo lagu saleynayo jiilka labaad.Tijaabadu uma baahna inay ku darto wax enzymes ah, RNA si fudud ayaa loo heli karaa laba tiir oo keliya.

Tiirka 1: Tiirka Nadiifinta DNA si loo nadiifiyo DNA

Tiirka 2: Tiirka-RNA-kaliya ayaa si gaar ah u soo kabanaya oo soo kabanaya RNA

Hawlgalka fudud ee noocan oo kale ah ma aha oo kaliya inuu badbaadiyo wakhti, laakiin sidoo kale wuxuu fududeeyaa hawlgalka tijaabooyinka hoose!

Alaabta matale: QIAGEN RNeasy Plus iyoForegeneRNA sifaynta alaabta taxanaha ah

(RNeasy Plus)

(Foregene'sQalabka Go'doominta Dhirta ee RNA

Aamusnaan kuugu sheeg!Qalabka RNA ee Foregene waxa kale oo uu soo bandhigayaa qalab aad u fudud in la isticmaalo RNase Eraser, kaas oo gabi ahaanba ka saari kara 400μg ee RNase dusha adag 1 daqiiqo gudaheed, iyada oo aan lagu darin wax walxo sun ah jidhka bini'aadamka!

Sawir1: shaxda isbarbardhigga habka hawlgalka

Alaabooyinka Foregene waxay ku dhammaystiri karaan soo saarista RNA laba heesood oo keliya, ha ka dhigin kuwa kale inay ka masayraan horumarka tijaabadaada!

Sawir2:Enatiijooyin tijaabo ah shaxda isbarbardhigga

Jiilka kowaad (1: TRIzol)

Jiilka labaad (2: RNeasy ee QIAGEN)

Jiilka saddexaad (3: QIAGEN's RNeasy lagu daray, 4: Foregene)

Sawir3: garaafka qPCR (garaaf qalooca kor u qaadista)

Jiilka kowaad (cagaaran macnaheedu waa TRIzol -, casaan macnaheedu waa TRIzol +)

Jiilka labaad (buluug macnaheedu waa QIAGEN RNeasy-, casaan macnaheedu waa QIAGEN RNeasy +)

Jiilka saddexaad (buluug macnaheedu waa QIAGEN RNeasy oo lagu daray, cagaar macnaheedu waa Foregene)

(Fiiro gaar ah: garaafka qPCR "-" wuxuu muujinayaa in DNA-da aan lagu darin oo aan la saarin wasakheynta DNA-da. Sidaa darteed, CT-ga ayaa sarreeya oo natiijadu maaha mid sax ah; "+" waxay muujinaysaa in DNA lagu daray oo wasakhda DNA laga saaray, natiijadu waa sax)

Casriyeyn kasta oo farsamo macnaheedu waa in hawlgalka tijaabada uu ku kobcayo jihada fudud oo ka badan isticmaale-saaxiibtinimo.

Foregene ujeedadeedu tahay inay caawiso cilmi-baarayaal badan oo saynis ah inay badbaadiyaan waqtiga tijaabada iyo kharashka, helaan natiijooyin tijaabo ah oo tayo sare leh, iyo si wadajir ah u abuuraan guulo cilmi baaris oo qiimo leh.