RNAprep Pure Cell/Bacteria Kit Featured Image

RNAprep Isisele esinyulu / iBacteria Kit

Ukucoca i-RNA ekumgangatho ophezulu kwiiseli nakwibhaktheriya.

I-RNAprep Iselfowuni / iBacteria Kit esulungekileyo ibonelela ngendlela ekhawulezayo, elula kwaye engabizi mali ininzi yokuhlanjululwa kwe-RNA iyonke kwiiseli ezikhulisiweyo kunye neesampulu zebhaktiriya ngokusebenzisa ikholamu efanelekileyo yokujikeleza kunye nenkqubo eyingqayizivele yesixhobo. Ikhithi ibandakanya i-RNase-Free Spin Column CR3 yokucoca i-RNA ekumgangatho ophezulu ngokusebenzisa itekhnoloji ye-silica-membrane. Umgangatho opheleleyo we-RNA unokufunyanwa kwimizuzu engama-30 ukuya kwengama-40 ngokusulungeka okuphezulu kwaye akukho simahla kwiprotein kunye ne-genomic DNA ungcoliseko.

Ikati. Hayi	Ubungakanani bokuPakisha
4992235	50 preps

Thumela i-imeyile kuthi

Iinkcukacha Product

Umzekelo wovavanyo

FAQ

Iimpawu zemveliso

Iimbonakalo

Buff Izilondolozi ezilungiselelwe oko kunye neeprotokholi zeeseli ezikhulisiweyo kunye neesampulu zebhaktiriya ziyenza inkqubo ibe lula kwaye ifaneleke.
■ I-DNase eyahlukileyo ndiyacutha ukungcoliseka kwe-genomic DNA.
■ Iikholamu ezikhethekileyo ze-RNase-Free Filtration Columns CS zishenxisa olunye ungcoliseko.
■ Ukucoceka okuphezulu kunye nokulungele ukusetyenziswa kwe-RNA kufanelekile kwizicelo ezibuthathaka ezantsi.
■ Akukho kucolwa kwephenol / chloroform, akukho LiCl kunye nemvula ethanol, akukho CsCl gradient centrifugation iyafuneka, eyenza inkqubo ikhuseleke kwaye ithembeke.

Izicelo

■ I-RT-PCR.
■ Iblukhwe laseMantla, iBala eliMachaphaza.
■ I-PCR yokwenene.
■ Uhlalutyo lwe-Chip.
■ Ukuvavanywa kwePolyA, kuguqulelo lwe-vitro, uhlalutyo lokhuselo lwe-RNase kunye nokwenziwa kwemolekyuli.

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

Egqithileyo I-TRNzol yendalo iphela

Okulandelayo: I-RNAprep Kit efanelekileyo yeeTishu

	Isixhobo: Iiseli zeJurkat zoLuntu (1 × 10⁶ ) Indlela: I-RNA iyonke yeeSeli zeJukat zoLuntu zahlukaniswa zisebenzisa i-RNAprep Pure Cell / Bacteria Kit. Iziphumo: Nceda ubone lo mfanekiso ungasentla we-agarose gel electrophoresis. I-2-4 μl ye-50 μl eluates ilayishwe ngomgaqo ngamnye. I-electrophoresis yaqhutywa nge-6 V / cm kwimizuzu engama-30 kwi-1% yejeli ye-agarose.
	Izinto eziphathekayo: TOP10 E. coli (1 × 10⁸) Indlela: I-RNA iyonke ye-TOP10 E. coli yahlulwa yodwa kusetyenziswa i-RNAprep Pure Cell / Bacteria Kit. Iziphumo: Nceda ubone lo mfanekiso ungasentla we-agarose gel electrophoresis. I-2-4 μl ye-50 μl eluates ilayishwe ngomgaqo ngamnye. I-electrophoresis yaqhutywa nge-6 V / cm kwimizuzu engama-30 kwi-1% yejeli 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).

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