Weli miyaad isticmaashaa Trizol si aad u soo saarto RNA?
Allahayow, caafimaadkaaga ha iga welwelin!
Trizol waxay ku dhowdahay in la takooro, ha ii sheegin inaad weli mugdi ku jirto?

Qaybta ugu muhiimsan ee reagent Trizol waa phenol, iyo reagents organic sida chloroform ayaa sidoo kale lagu daray hawlgalka tijaabada ah.Soo-gaadhista muddada-dheer waxay dhaawac u geysan doontaa caafimaadkaaga, gaar ahaan taabashada tooska ah ee maqaarka waxay sababi kartaa gubasho kiimiko oo daran iyo nabarro joogto ah.

Xaaladaha caadiga ah, wada-hawlgalayaasha shaybaarku waxay yareyn karaan khataraha iyaga oo si buuxda isu hubeeya (xiran jaakadaha shaybaadhka, galoofyada iyo maaskaro).

Talada aan ku siinayaana waa: fadlan si toos ah u beddel Trizol si aad uga fogaato waxyeellada ka imanaysa isha.

1. Shayga ugu horreeya ee soo saarista RNA
Soo baxay 1987 (Trizol reagents)

faa'iido:
Soosaar dhammaystiran oo dheeraad ah

Khasaaraha:
Reageniyeyaasha sunta ah sida guanidine isothiocyanate-phenol-chloroform ayaa loo baahan yahay;
Qalliin dhib badan;
Loo baahan yahay in la isticmaalo DNA-da si looga saaro wasakheynta DNA-da

2.Waxyaabaha jiilka labaad ee soo saarista RNA
Soo bax 1994 (QIAGEN RNeasy iyo inta badan alaabta gudaha)

faa'iido:
Habka centrifugal tiirarka ayaa la qaatay, kaas oo fudud in la shaqeeyo;

Khasaaro:
Waxa loo baahan yahay in la centrifuge si meesha looga saaro haraaga unugyada
Loo baahan yahay in la isticmaalo DNA-da si looga saaro wasakheynta DNA-da

3.Wax soo saarka jiilka saddexaad ee soo saarista RNA
Soo baxay 2005 (QIAGEN RNeasy Plus iyo ForegeneAlaabooyinka taxanaha ah ee nadiifinta RNA)

faa'iido:
Si toos ah u isticmaal tiirka saarista DNA si aad u tirtirto DNA-da iyo qashinka unugyada isku mar, badbaadinta wakhtiga iyo dadaalka;
Badbaadsan oo ku habboon, looma baahna in lagu daro tiirka shaandhada iyo DNA-da si si fudud looga saaro wasakhda loogana fogaado wasakhaynta DNA-da

Aynu eegno kala duwanaanshaha hawlgalka ee u dhexeeya badeecada jiilka koowaad (Trizol), badeecada jiilka labaad (RNeasy) iyo badeecada jiilka saddexaad (Foregene) ee soo saarista RNA:

Hawlgalka shaxanka waa cad yahay: sheyga saddexaad ee jiilka ayaa leh faa'iidooyin muuqda

Kaliya saddex kelmadood oo lagu tilmaamo: fudud!Degdeg ah!Badbaadada

Shaxda PK ee natiijooyinka tijaabadawaayojiilka kowaad, jiilka labaad iyo jiilka saddexaad ee soo saarista RNA:

1. Electrophoretogram

Jiilka koowaad (1: Trizol)
Jiilka labaad (2:QIAGEN RNeasy)
Jiilka saddexaad (3: QIAGEN RNeasy oo lagu daray, 4: Foregene)

Waa maxay nooca badeecada fiican, waxaad ogaan doontaa marka la barbardhigo
RNA ayaa la soo saaray iyadoo la isticmaalayo qalabka Foregene
Ma aha oo kaliya feejignaan sare laakiin sidoo kale daacadnimo wanaagsan

2.qPCR garaaf (garaaf qalooca kor u qaadida)

Jiilkii kowaad (Trizol)

Jiilkii labaad (sky bluemacneheedu waaQIAGEN RNeasy-, casaanmacneheedu waaQIAGEN RNeasy +)

Jiilka saddexaad (buluug macnaheedu waa Foregene, cagaar macnaheedu waa QIAGEN RNeasy plus)

(Fiiro gaar ah: Jiilka labaad ee QIAGEN RNeasy ee qPCR garaaf macnaheedu waa in qalabku aanu ku darin DNA-ga oo aanu ka saarin wasakhaynta DNA-da. Sidaa darteed, CT-gu wuu sarreeyaa natiijaduna maaha mid sax ah; QIAGEN RNeasy + macnaheedu waa in DNA lagu daray oo wasakheynta DNA laga saaray, natiijaduna waa sax)

Haddi ay tahay electrophoretogram ama garaaf qalooca PCR tirade, tayada alaabta Foregene kama liidato alaabta dibada laga keeno.