RNAclean Kit

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).