RNAsimple Yakazara RNA Kit
Zvimiro
■ Iyo yakachena-kuchena yakagadzirira-kushandisa RNA inokodzera kune inonzwisisika yekudzika mafomu.
■ Wide mafomu: Iyo yakanatswa RNA inogona kuiswa kune akasiyana ekuyedza sampuli.
The Kuedza kwacho kunogona kupedzwa muawa rimwe chete nemabasa ari nyore.
Zvishandiso
■ RT-PCR
■ Northern Blot, Dot Blot.
■ Real-Nguva PCR
■ PolyA Kuongorora, in vitro dudziro, RNase yekudzivirira ongororo, mamorekuru kuumbana.
Zvese zvigadzirwa zvinogona kuve zvakagadziriswa ODM / OEM. Kuti uwane mamwe mashoko,ndokumbira ubaye Customized Service (ODM / OEM)
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Chinyorwa: 20 mg rat rateti spleen tishu Maitiro: Iyo yakazara RNA yemakonzo spleen tishu yaive yakasarudzika ichishandisa iyo RNAsimple Yese RNA Kit. Mhedzisiro: Ndokumbira kuti uone iri pamusoro agarose gel electrophoresis pikicha. 2-4 μl e100 μl eluates akatakurwa pamugwagwa. Iyo electrophoresis yakaitwa pa6 V / cm ye30 min pa1% agarose. |
A-1 Cell lysis kana homogenization haina kukwana
- Deredza kushandiswa kwesampula, wedzera huwandu hwerudzi rwe lysis, wedzera homogenization uye lysis nguva.
A-2 Mhando yemuenzaniso yakanyanya kukura
--Deredza huwandu hwemuenzaniso wakashandiswa kana kuwedzera huwandu hwerisiti bhaisikopo.
A-1 Isina kukwana sero lysis kana homogenization
- Deredza kushandiswa kwesampula, wedzera huwandu hwerudzi rwe lysis, wedzera homogenization uye lysis nguva.
A-2 Mhando yemuenzaniso yakanyanya kukura
- Ndokumbira utarise kunonyanyisa kugadzira chinzvimbo.
A-3 RNA haina kukwidziridzwa zvizere kubva pambiru
- Mushure mekuwedzera RNase-Yemahara mvura, siya iyo kwemaminetsi mashoma pamberi pe centrifuging.
A-4 Ethanol iri mudenga
- Mushure mekuchenesa, centrifuge zvakare uye bvisa iyo yekugezera bhafa zvakanyanya sezvinobvira.
A-5 Cell tsika yepakati haina kubviswa zvachose
- Paunenge uchiunganidza maseru, ndapota ita chokwadi chekubvisa iyo tsika yepakati zvakanyanya sezvinobvira.
A-6 Masero akachengetwa muRNAstore haana kunyatsoita centrifuged
- RNAstore density yakakura kupfuura avhareji sero tsika svikiro; saka simba re centrifugal rinofanira kuwedzerwa. Inokurudzirwa kucentrifuge pa3000x g.
A-7 Yakadzika RNA zvemukati uye kuwanda mune muyenzaniso
- Shandisa yakanaka sampuro kuona kana yakaderera-goho inokonzerwa nesampuli.
A-1 Chinyorwa hachisi chitsva
- Nyowani nyowani inofanirwa kuchengetwa muiriidhi senitrogen nekukasira kana nekukurumidza kuiswa muRNAstore reagent kuve nechokwadi chekudzora mhedzisiro.
A-2 Mhando yemuenzaniso yakanyanya kukura
- Deredza sampuro yemari.
A-3 RNase kusvibiswan
- Kunyangwe iyo buffer yakapihwa mune kit haina RNase, zviri nyore kusvibisa RNase panguva yekubvisa uye inofanira kubatwa nehanya.
A-4 Electrophoresis kusvibiswa
- Tsiva iyo electrophoresis buffer uye ita shuwa kuti zvinoshandiswa uye Kurodha Buffer hazvina kusvibiswa neRNase.
A-5 Kunyanya kurodha electrophoresis
--Deredza huwandu hwesampuli kurodha, kurodha kwetsime rega rega hakufanirwe kupfuura 2 μg.
A-1 Mhando yemuenzaniso yakanyanya kukura
- Deredza sampuro yemari.
A-2 Mimwe mienzaniso ine yakakwira DNA yezviri mukati uye inogona kurapwa neDNase.
- Ita RNase-Yemahara DNase kurapwa kune yakawanikwa RNA mhinduro, uye iyo RNA inogona kushandiswa zvakananga kuzviedza zvinotevera mushure mekurapwa, kana inogona kuvhenekerwa neRNA yekuchenesa makiti.
Kune magirazi egirazi, akabikwa pa150 ° C kwe4 h. Pamidziyo yepurasitiki, yakanyudzwa mu0.5 M NaOH kwemaminetsi gumi, ndokunyatsogezwa nemvura isina RNase uye wozadza kuti ubvise zvachose RNase. Iwo ma reagents kana mhinduro dzinoshandiswa mukuyedza, kunyanya mvura, dzinofanirwa kunge dzisina RNase. Shandisa RNase-yemahara mvura kune ese reagent gadziriro (wedzera mvura kune yakachena girazi bhodhoro, wedzera DEPC kune yekupedzisira fungidziro ye0.1% (V / V), zunza husiku uye autoclave).
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