RNAprep Koyera yaying'ono zida

Kuyeretsa RNA yokwanira kuchokera kuzinthu zazing'ono kapena maselo.

RNAprep Pure Micro Kit imagwiritsa ntchito gawo loyendetsa bwino kwambiri, la nucleic acid-centrifugal adsorption ndi chida chapadera chotengera RNA yonse pamitundu mitundu ingapo yama microsamples. Chida ichi chimaphatikizapo Chonyamulira RNA, chomwe chimatha kupeza ma nucleic acid mosavuta m'dongosolo. Kutulutsa kwa RNA ndi chida ichi ndikosavuta, mwachangu komanso kuberekanso ndi zokolola zambiri. Zomwe zimachitika zitha kumaliza mkati mwa mphindi 30-40. Chikwamacho chimatha kuchotsa RNA yonse yomwe ili <200 nt (5.8S rRNA, 5S rRNA ndi tRNAs, ndi zina zambiri), pomwe imalimbikitsa, yeretsani ndikuyeretsa RNA yonse yomwe ili> 200 nt. RNA yonse yotulutsidwa ndi yoyera kwambiri komanso yopanda DNA komanso kuipitsidwa kwa mapuloteni.

Mphaka. Ayi	Kukulitsa Kukula
4992859	50 preps

Tumizani imelo kwa ife

Mankhwala Mwatsatanetsatane

Chitsanzo Cha Zoyesera

FAQ

Zogulitsa

Mawonekedwe

■ Kutha kuyeretsa RNA yapamwamba kwambiri pamitundu yotsata, monga tinthu tating'onoting'ono tating'onoting'ono, minofu yolimba ndi maselo.
■ DNase Wapadera I amachepetsa kuipitsidwa kwa DNA.
■ RNA yoyera kwambiri komanso yogwiritsidwa ntchito ndiyabwino kuchitira tcheru kumapeto.
■ Palibe chochotsa phenol / chloroform, palibe LiCl ndi mpweya wa ethanol, palibe CsCl gradient centrifugation yomwe ikufunika, zomwe zimapangitsa kuti ntchitoyi ikhale yotetezeka komanso yodalirika.

Mapulogalamu

■ RT-PCR.
■ Northern Blot, Dot Blot.
■ PCR Yeniyeni.
■ Kusanthula kwa Chip.
■ Kuwunika kwa PolyA, kumasulira kwa ma vitro, kusanthula kwa RNase ndi kupangika kwa ma molekyulu.

Zogulitsa zonse zimatha kusinthidwa kukhala ODM / OEM. Zambiri,chonde dinani Makonda Service (ODM / OEM)

Previous: Zamgululi RNAsimple Chiwerengero cha RNA Kit

Ena: TRNzol Universal Reagent

RNA yonse ya 1 × 10⁶, 1 × 10⁵, 1 × 10⁴, 1 × 10³, 1 × 10², Maselo 10 a Hela adachotsedwa pogwiritsa ntchito RNAprep Pure Micro Kit. RT-qPCR idachitidwa pogwiritsa ntchito Quant qRT-PCR (SYBR Green) Kit ya TIANGEN.

Q: Kutsekeka kwamagawo

A-1 Cell lysis kapena homogenization sikokwanira

---- Kuchepetsa ntchito nyemba, kuonjezera kuchuluka kwa gawo lotetezera lysis, kuonjezera homogenization ndi nthawi lysis.

A-2 Zitsanzo kuchuluka ndi waukulu kwambiri

---- Kuchepetsa kuchuluka kwa nyemba zomwe mukugwiritsa ntchito kapena kuonjezera kuchuluka kwa lysis buffer.

Q: Zokolola zochepa za RNA

A-1 Kusakwanira kwa cell lysis kapena homogenization

---- Kuchepetsa ntchito nyemba, kuonjezera kuchuluka kwa gawo lotetezera lysis, kuonjezera homogenization ndi nthawi lysis.

A-2 Zitsanzo kuchuluka ndi waukulu kwambiri

- Chonde onani kuthekera kwakukulu kokonza.

A-3 RNA siyosankhidwa kwathunthu kuchokera pagawo

---- Mukatha kuwonjezera madzi opanda RNase, siyani kwa mphindi zochepa centrifuging.

A-4 Ethanol mumlengalenga

- Pambuyo kutsukidwa, centrifuge kachiwiri ndikuchotsa chotsuka chotsuka momwe zingathere.

A-5 Cell chikhalidwe sing'anga sichichotsedweratu

- Mukamasonkhanitsa maselo, chonde onetsetsani kuti muchotse sing'anga momwe mungathere.

A-6 Maselo osungidwa mu RNAstore sakhala otsekemera kwenikweni

---- RNAstore osalimba ndi wamkulu kuposa sing'anga wamba wamaselo; kotero mphamvu ya centrifugal iyenera kukulitsidwa. Akuti centrifuge pa 3000x g.

Zolemba za A-7 Low RNA komanso kuchuluka kwake mchitsanzo

- Gwiritsani ntchito chitsanzo chabwino kuti muwone ngati zokolola zochepa zimayambitsidwa ndi chitsanzocho.

Q: Kuwonongeka kwa RNA

A-1 Nkhaniyi siyatsopano

- Minyewa yatsopano imayenera kusungidwa mu nayitrogeni wamadzi nthawi yomweyo kapena kuyiyika mu reagent ya RNAstore kuti zitsimikizidwe kuti zimachokera.

A-2 Zitsanzo kuchuluka ndi waukulu kwambiri

---- Kuchepetsa nyemba kuchuluka.

A-3 RNase kuipitsidwan

- Ngakhale gawo lotetezedwa mulibe RNase, ndikosavuta kuipitsa RNase panthawi yazochulukirapo ndipo iyenera kusamalidwa.

Kuwononga kwa A-4 Electrophoresis

---- Sinthanitsani cholumikizira cha electrophoresis ndikuwonetsetsa kuti zofunikira ndi Kutsegula Buffer zilibe vuto la RNase.

A-5 Kutsitsa kwambiri kwa electrophoresis

---- Kuchepetsa kuchuluka kwa potsegula, kutsitsa kwa chitsime chilichonse sikuyenera kupitirira 2 μg.

Q: kuipitsidwa kwa DNA

A-1 Zitsanzo kuchuluka ndi waukulu kwambiri

---- Kuchepetsa nyemba kuchuluka.

A-2 Zitsanzo zina zimakhala ndi DNA yambiri ndipo amatha kuchiritsidwa ndi DNase.

- Chitani chithandizo cha RNase-Free DNase ku yankho la RNA, ndipo RNA itha kugwiritsidwa ntchito mwachindunji poyeserera pambuyo pothandizidwa, kapena itha kuyeretsedwa ndi zida zoyeserera za RNA.

Q: Kodi mungachotse bwanji RNase pazomwe mungagwiritse ntchito poyesera ndi magalasi?

Zamagalasi, zophikidwa pa 150 ° C kwa 4 h. Pazitsulo zapulasitiki, zomizidwa mu 0,5 M NaOH kwa mphindi 10, kenako kutsukidwa bwino ndi madzi opanda RNase kenako ndikuwotcha kuti muchotse RNase. Ma reagents kapena mayankho omwe amagwiritsidwa ntchito poyeserera, makamaka madzi, ayenera kukhala opanda RNase. Gwiritsani ntchito madzi opanda RNase pamakonzedwe onse a reagent (onjezerani madzi mu botolo loyera lagalasi, onjezerani DEPC pamlingo womaliza wa 0,1% (V / V), gwedezani usiku umodzi ndi autoclave).

Lembani uthenga wanu apa ndi kutumiza kwa ife