I-RNAsimple Total RNA Kit

Ngokukhishwa okuphelele okuphelele kwe-RNA kusetshenziswa ikholomu ye-centrifugal esetshenziswa kakhulu.

I-RNAsimple Total RNA Kit isebenzisa indlela entsha yokukhipha i-RNA ngokususelwa ku-guanidinium isothiocyanate / phenol method. I-Buffer RZ eyenziwe ngokukhethekile yi-TIANGEN ingasusa i-genomic DNA namaprotheni kumaseli ngokushesha nangokuphumelelayo, okwenza i-RNA etholakalayo ibe msulwa kakhulu futhi izinzile.
Le kit isetshenziselwa ukuhlukanisa i-RNA ephelele egazini, kumaseli ezilwane, izicubu nezicubu zezitshalo. Ikholomu ngayinye ye-spin ingacubungula izicubu ezingama-50-100 mg noma i-5 × 10⁶amaseli ngasikhathi sinye futhi angacubungula inani elikhulu lamasampuli ahlukene ngasikhathi sinye. Ukusabela kungaqedwa kungaphansi kwehora elilodwa, futhi ingqikithi ye-RNA ekhishwe inesivuno esiphezulu, ubumsulwa obungcono, ngaphandle kokungcola kwe-DNA kanye namaprotheni, futhi ingasetshenziswa ekuhlolweni okuhlukahlukene okwehla komfula.

Ikati. Cha	Ukupakisha Usayizi
4992858	Ama-preps angama-50

Thumela i-imeyili kithi

Imininingwane yomkhiqizo

Ukuhamba komsebenzi

Isibonelo sokuhlola

Imibuzo evame ukubuzwa

Amathegi womkhiqizo

Izici

■ Ukuhlanzeka okuphezulu okusetshenziselwa ukusetshenziswa kwe-RNA kufanelekile ekusetshenzisweni okubucayi komfula.
■ Izicelo ezibanzi: I-RNA ehlanziwe ingasetshenziswa kumasampula wokuhlola ahlukahlukene.
■ Ukuhlolwa kungaphothulwa ngehora eli-1 ngokusebenza okulula.

Izicelo

■ I-RT-PCR
■ I-Northern Blot, iDot Blot.
■ I-PCR Yesikhathi Sangempela
■ Ukuhlolwa kwe-PolyA, ukuhumusha kwe-in vitro, ukuhlaziywa kwe-RNase yokuvikela, ukuhlanganiswa kwamangqamuzana.

Yonke imikhiqizo ingenziwa ngezifiso i-ODM / OEM. Ngemininingwane,sicela uchofoze i-Customized Service (ODM / OEM)

Langaphambilini I-RNAclean Kit

Olandelayo: I-RNAprep Pure Micro Kit

Okubalulekile: Izicubu zomgogodla wobuchopho obungu-20 mg
Indlela: Ingqikithi ye-RNA yezicubu zomzimba wamagundane yahlukaniswa kusetshenziswa i-RNAsimple Total RNA Kit.
Imiphumela: Sicela ubheke isithombe esingenhla se-agarose gel electrophoresis. I-2-4 μl yama-100 μl eluates alayishwe kulayini ngamunye. I-electrophoresis yenziwa ngo-6 V / cm ngemizuzu engama-30 ku-1% agarose.

Q: Ukuvinjelwa kwekholomu

I-A-1 Cell lysis noma i-homogenization ayanele

- Nciphisa ukusetshenziswa kwesampula, khulisa inani le-lysis buffer, khulisa i-homogenization nesikhathi se-lysis.

Inani elingu-A-2 lesampuli likhulu kakhulu

---- Yehlisa inani lesampula elisetshenzisiwe noma wandise inani le-lysis buffer.

Q: Isivuno se-RNA esiphansi

I-A-1 Inganele i-lysis yeseli noma i-homogenization

- Nciphisa ukusetshenziswa kwesampula, khulisa inani le-lysis buffer, khulisa i-homogenization nesikhathi se-lysis.

Inani elingu-A-2 lesampuli likhulu kakhulu

---- Sicela ubheke amandla wokucubungula aphezulu.

I-A-3 RNA ayikhishwa ngokuphelele kukholamu

---- Ngemuva kokungeza amanzi e-RNase-Free, yiyeke imizuzu embalwa ngaphambi kwe-centrifuging.

I-A-4 Ethanol ku-eluent

---- Ngemuva kokuhlanza, i-centrifuge futhi bese ususa i-buffer yokuwasha ngangokunokwenzeka.

I-A-5 Cell culture medium ayisuswanga ngokuphelele

- Lapho uqoqa amaseli, sicela uqiniseke ukuthi ususa okuphakathi kwesiko ngangokunokwenzeka.

A-6 Amaseli agcinwe ku-RNAstore awasebenzi kahle nge-centrifuged

---- Ukuxinana kwe-RNAstore kukhulu kunesilinganiso esiphakathi sesiko leseli; ngakho-ke amandla e-centrifugal kufanele akhuliswe. Kuphakanyiswa ku-centrifuge ku-3000x g.

Okuqukethwe kwe-A-7 okuphansi kwe-RNA nobuningi kusampula

---- Sebenzisa isampula elihle ukuthola ukuthi isivuno esiphansi sidalwa yisampula.

Q: Ukucekelwa phansi kwe-RNA

A-1 Indaba ayisiyintsha

Izicubu ezintsha kufanele zigcinwe ku-nitrogen ewuketshezi ngokushesha noma ngokushesha zifakwe kwi-reagent ye-RNAstore ukuqinisekisa umphumela wokukhishwa.

Inani elingu-A-2 lesampuli likhulu kakhulu

---- Yehlisa inani lesampula.

I-A-3 RNase contamination

- Yize i-buffer enikezwe kukhithi ingenayo i-RNase, kulula ukungcolisa i-RNase ngesikhathi senqubo yokukhipha futhi kufanele iphathwe ngokunakekela.

Ukungcola kwe-A-4 Electrophoresis

---- Faka esikhundleni se-electrophoresis buffer futhi uqiniseke ukuthi izinto ezisetshenziswayo ne-Loading Buffer azinakho ukungcoliswa yi-RNase.

A-5 Kulayishwa kakhulu i-electrophoresis

---- Yehlisa inani lokulayishwa kwesampula, ukulayishwa komthombo ngamunye akufanele kudlule ku-2 μg.

Umbuzo: Ukungcola kwe-DNA

Inani elingu-1 lesampuli likhulu kakhulu

---- Yehlisa inani lesampula.

A-2 Amanye amasampula anokuqukethwe okuphezulu kwe-DNA futhi angalashwa nge-DNase.

---- Enza ukwelashwa kwe-RNase-Free DNase kusixazululo esitholakele se-RNA, futhi i-RNA ingasetshenziswa ngqo ekuhlolweni okulandelayo ngemuva kokwelashwa, noma ingaqhubeka ihlanzwe ngamakhithi wokuhlanzwa we-RNA.

Q: Ungayisusa kanjani i-RNase kokudliwayo kokuhlola kanye nama-glasswares?

Okwezingilazi, kubhakwe ku-150 ° C ngehora elingu-4. Kwiziqukathi zepulasitiki, ucwiliswe ku-0.5 M NaOH ngemizuzu eyi-10, bese ugeza kahle ngamanzi angenawo i-RNase bese uvala inzalo ukususa i-RNase ngokuphelele. Ama-reagents noma izixazululo ezisetshenziswe ekuhlolweni, ikakhulukazi amanzi, kumele zingabi ne-RNase. Sebenzisa amanzi angenawo ama-RNase kuwo wonke amalungiselelo e-reagent (engeza amanzi ebhodleleni lengilazi elihlanzekile, engeza i-DEPC ekugxileni kokugcina okungu-0.1% (V / V), unyakazise ubusuku kanye ne-autoclave).

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