RNAsimple Iyonke iRNA Kit

Ukukhutshwa okuphezulu okupheleleyo kwe-RNA kusetyenziswa ikholamu esetyenziswa ngokubanzi.

I-RNAsimple iyonke ye-RNA Kit yamkela indlela entsha yokukhupha i-RNA esekwe kwi-guanidinium isothiocyanate / phenol indlela. I-Buffer RZ eyilelwe ngokukodwa yi-TIANGEN ingasusa i-genomic DNA kunye neeproteni ezivela kwiiseli ngokukhawuleza nangokufanelekileyo, zisenza i-RNA efumanekayo ibe nyulu kwaye izinzile.
Le khithi isetyenziselwa ukwahlula i-RNA iyonke kwigazi, iiseli zezilwanyana, izicwili kunye nezicubu zezityalo. Ikholamu nganye yokujikeleza inokuqhubekeka izicubu ze-50-100 mg okanye i-5 × 106iiseli ngexesha kwaye zinokuqhubekeka inani elikhulu leesampulu ezahlukeneyo ngaxeshanye. Impendulo inokugqitywa kungaphelanga iyure enye, kwaye iyonke i-RNA ekhutshiweyo inesivuno esiphezulu, ubunyulu obungcono, ngaphandle kwe-DNA kunye neprotein yongcoliseko, kwaye inokusetyenziswa kuvavanyo olusezantsi.

Ikati. Hayi Ubungakanani bokuPakisha
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Iinkcukacha Product

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Umzekelo wovavanyo

FAQ

Iimpawu zemveliso

Iimbonakalo

■ Ukucoceka okulungele ukusetyenziswa kwe-RNA kufanelekile kwizicelo ezibuthathaka ezantsi.
■ Ukusetyenziswa okubanzi: I-RNA ecociweyo inokusetyenziswa kwiisampulu ezahlukeneyo zovavanyo.
The Uvavanyo lungagqitywa ngeyure e-1 ngemisebenzi elula.

Izicelo

■ I-RT-PCR
■ Iblukhwe laseMantla, iBala eliMachaphaza.
■ I-PCR yokwenene
■ Ukuvavanywa kwePolyA, kwi-vitro translation, uhlalutyo lokhuselo lwe-RNase, ukwenziwa kwemolekyuli.

Zonke iimveliso zingenziwa ngokwezifiso kwi-ODM / OEM. Iinkcukacha,nceda ucofe iNkonzo eChongiweyo (i-ODM / i-OEM)


  • Egqithileyo
  • Okulandelayo:

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    Workflow

    Experimental Example Isixhobo: 20 mg rat spleen tissue
    Indlela: I-RNA iyonke yeethisisi zethayile yaza yahlukaniswa kusetyenziswa iRNAsimple Iyonke iRNA Kit.
    Iziphumo: Nceda ubone lo mfanekiso ungasentla we-agarose gel electrophoresis. I-2-4 μl ye-100 μl eluates ilayishwe ngomgaqo ngamnye. I-electrophoresis yenziwa nge-6 V / cm kwimizuzu engama-30 kwi-1% ye-agarose.
    Q: Ikholamu yokuvalela

    I-A-1 Cell lysis okanye i-homogenization ayonelanga

    Ukunciphisa ukusetyenziswa kwesampula, ukwandisa inani le-buffer ye-lysis, ukwandisa i-homogenization kunye nexesha le-lysis.

    Isixa sesampulu esingu-A-2 sikhulu kakhulu

    ---- Ukunciphisa inani lesampuli esetyenzisiweyo okanye ukwandisa inani le-lysis buffer.

    Q: Isivuno seRNA esisezantsi

    A-1 I-lysis yeseli engonelanga okanye i-homogenization

    Ukunciphisa ukusetyenziswa kwesampula, ukwandisa inani le-buffer ye-lysis, ukwandisa i-homogenization kunye nexesha le-lysis.

    Isixa sesampulu esingu-A-2 sikhulu kakhulu

    - Nceda ubhekise kubungakanani bokusebenza.

    I-A-3 RNA ayikhutshiwe ngokupheleleyo kwikholamu

    --Ngasemva kokongeza amanzi asimahla e-RNase, yishiye imizuzu embalwa ngaphambi kokufaka isenti.

    I-A-4 Ethanol elufeni

    - Emva kokucoca, icentrifuge kwakhona kwaye ususe isikhuseli sokuhlamba kangangoko.

    I-A-5 yesiko lenkcubeko yeseli ayisuswanga ngokupheleleyo

    - Xa uqokelela iiseli, nceda uqiniseke ukuba ususa indawo yenkcubeko kangangoko kunokwenzeka.

    A-6 Iiseli ezigcinwe kwiRNAstore azisebenzi kakuhle kwisenti

    --Uxinano lweRNAstore lukhulu kunophakathi ophakathi kwenkcubeko yeseli; ke amandla e-centrifugal kufuneka anyuswe. Kuyacetyiswa kwi-centrifuge kwi-3000x g.

    Umxholo we-A-7 Low RNA kunye nobuninzi kwisampulu

    --Sebenzisa isampulu elungileyo ukumisela ukuba ngaba isivuno esisezantsi sibangelwa sisampulu.

    Q: Ukuthotywa kweRNA

    A-1 Imathiriyeli ayisiyonto intsha

    Izicubu ezintsha kufuneka zigcinwe kwi-nitrojeni engamanzi kwangoko okanye kwangoko ibekwe kwi-RNAstore reagent ukuqinisekisa ukusebenza kokukhutshwa.

    Isixa sesampulu esingu-A-2 sikhulu kakhulu

    ---- Ukunciphisa inani lesampula.

    A-3 RNase ukungcolan

    - Nangona i-buffer ebonelelwe kwikhithi ingenayo i-RNase, kulula ukungcolisa i-RNase ngexesha lenkqubo yokukhupha kwaye kufuneka iphathwe ngononophelo.

    Ungcoliseko lwe-4-Electrophoresis

    ---- Faka endaweni ye-electrophoresis buffer kwaye uqiniseke ukuba izinto ezisetyenziswayo kunye nokuLayisha iBuffer zikhululekile kungcoliseko lweRNase.

    A-5 Ukulayisha kakhulu i-electrophoresis

    ---- Ukunciphisa inani lokulayisha isampula, ukulayishwa kwequla ngalinye akufuneki kudlule kwi-2 μg.

    Umbuzo: Ungcoliseko lwe-DNA

    A-1 Isixa sesampulu sikhulu kakhulu

    ---- Ukunciphisa inani lesampula.

    A-2 Ezinye iisampulu zinomxholo ophezulu we-DNA kwaye zinokunyangwa nge-DNase.

    - Yenza unyango lwe-RNase-Simahla ye-DNase kwisisombululo esifunyenweyo se-RNA, kwaye i-RNA inokusetyenziselwa ngokuthe ngqo kuvavanyo olulandelayo emva konyango, okanye inokucocwa ngakumbi ziikhithi zokucocwa zeRNA.

    Q: Uyisusa njani i-RNase kwizinto ezisetyenziswayo zovavanyo nakwiglasi?

    Ngeiglasi, kubhakwe kwi-150 ° C nge-4 h. Kwizikhongozeli zeplastiki, zibhaptizwe kwi-0.5 M NaOH kangange-10 min, emva koko zahlanjwa kakuhle ngamanzi angenasimahla e-RNase kwaye emva koko zibulale ukuze ususe i-RNase ngokupheleleyo. Izinto ezisetyenziswayo okanye isisombululo esisetyenzisiweyo kulingo, ngakumbi amanzi, kufuneka simahla kwi-RNase. Sebenzisa amanzi asimahla e-RNase kuwo onke amalungiselelo e-reagent (yongeza amanzi kwibhotile yeglasi ecocekileyo, yongeza i-DEPC kuxinzelelo lokugqibela lwe-0.1% (V / V), gungqisa ubusuku kunye ne-autoclave).

    Bhala umyalezo wakho apha uze uyithumele kuthi