Qalabka Tooska ah ee Unugga RT QPCR-SYBR GREEN I

Hordhac

◮ fudud oo waxtar lehTignoolajiyada Cell Direct RT, shaybaarka RNA waxaa lagu heli karaa 7 daqiiqo oo keliya.

Baahida muunada waa mid yar, maadaama ay yar tahay 10 unug ayaa la tijaabin karaa.

◮ Wax soo saarka sare: waxay si dhakhso ah u ogaan kartaa RNA unugyada lagu beeray 384, 96, 24, 12, 6-cells plates.

Tirtiraha DNA-da ayaa si dhakhso ah uga saari kara genome-yada la sii daayay, si weyn ayuu u yareeyaa saamaynta natiijooyinka tijaabada dambe ee xiga.

Habka RT-qPCR ee la hagaajiyay ayaa ka dhigaya labada-tallaabo ee RT-PCR qoraalka dib u dhigista mid hufan iyo PCR si gaar ah, oo aad u adkeysi badan u leh celiyaasha falcelinta RT-qPCR.

Faahfaahinta Alaabta

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Sharaxaada

Xirmadani waxay adeegsataa nidaam u gaar ah lysis-dhise kaas oo si dhakhso ah uga sii deyn kara RNA muunadaha unugyada dhaqameed ee falcelinta RT-qPCR, taas oo meesha ka saaraysa habka nadiifinta RNA waqti-qaadka iyo tacabka badan.Qaabka RNA waxa lagu heli karaa 7 daqiiqo oo kaliya.5× Toos ah RT Mix iyo 2× Toos ah qPCR Mix-SYBR reagents ay bixiso qalabku waxay si degdeg ah oo wax ku ool ah u heli karaan natiijooyinka PCR-ga dhabta ah.

5× Toos ah RT Mix iyo 2× qPCR Mix-SYBR toos ah waxay leeyihiin dulqaad adag oo ka hortag ah, iyo lysate ee shaybaarada waxaa loo isticmaali karaa qaab-dhismeedka RT-qPCR si toos ah.Qalabkani waxa uu ka kooban yahay RNA-ga gaarka ah ee xidhiidhka sare ee Foregene reverse transcriptase, iyo Hot D-Taq DNA polymerase, dNTPs, MgCl2, falcelinta falcelinta, PCR optimizer iyo stabilizer.

Tilmaamaha

200×20μl Rxns, 1000×20μl Rxns

Qaybaha kitka

Qaybta I

Buffer CL

Foregene Protease Plus II

Buffer ST

Qaybta II

Tirtiraha DNA

5× Toos ah RT Mix

2× Toos ah qPCR Mix-SYBR

50× ROX Tixraaca Midabka

RNase-free ddH2O

Tilmaamaha

Tilmaamaha & faa'iidooyinka

■ Fudud oo wax ku ool ah: tignoolajiyada Cell Direct RT, muunadaha RNA waxaa lagu heli karaa 7 daqiiqo oo keliya.

■ Baahida muunada waa yar tahay, maadaama ay yar tahay 10 unug ayaa la tijaabin karaa.

■ Soo saarista sare: waxay si dhakhso ah u ogaan kartaa RNA ee unugyada lagu beeray 384, 96, 24, 12, 6-cells.

■ Tirtiraha DNA-da si dhakhso ah ayuu u saari karaa genome-yada la sii daayay, si weyn ayuu u yareynayaa saameynta natiijooyinka tijaabada dambe.

■ Nidaamka RT iyo qPCR ee la hagaajiyay ayaa ka dhigaya laba-tallaabo dib-u-celinta RT-PCR mid hufan iyo PCR mid gaar ah, oo aad u adkeysi badan ka-hortagga falcelinta RT-qPCR.

Codsiga qalabka

Baaxadda codsiga: unugyada dhaqanka.

- RNA lagu sii daayay muunad lysis: kaliya lagu dabaqi karo qaabka RT-qPCR ee xirmadan.

-Qalabka waxa loo isticmaali karaa ujeedooyinka soo socda: falanqaynta muujinta hidde-sidaha, xaqiijinta saamaynta aamusinta hidde-sidaha ee siRNA, baadhista daroogada, iwm.

Jaantuska

Unug-toos ah-RT-qPCR-shaxanka

 

Kaydinta iyo nolosha shelf

Qaybta I ee xirmadan waa in lagu kaydiyaa 4℃;Qaybta II waa in lagu kaydiyaa at -20 ℃.

Foregene Protease Plus II waa in lagu kaydiyaa 4 ℃, ha ku barafaysan -20 ℃.

Reagent 2× Toos ah qPCR Mix-SYBR waa in lagu kaydiyaa -20℃ gudcurka;haddii si joogto ah loo isticmaalo, waxa kale oo lagu kaydin karaa 4℃ kaydinta muddada gaaban (la isticmaalo 10 maalmood gudahood).


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