Imiyoboro yo gusesengura ibibazo

The following is an analysis of the problems that might be encountered in the extraction of viral RNA. We wish it would be helpful to your experiment. In addition, for other experimental or technical problems other than operating instructions and problem analysis, we have dedicated technical support to help you. Contact us if you need at : 028-83361257or E-mail：Tech@foregene.com。

Nta RNA ishobora gukururwa cyangwa umusaruro wa acide nucleic ni muke

Mubisanzwe hariho ibintu byinshi bigira ingaruka kumikorere yo gukira, nka: icyitegererezo cya RNA, uburyo bwo gukora, ingano ya elution, nibindi .。

Isesengura ry'impamvu zisanzwe ：

1.Kora ubwogero cyangwa ubushyuhe buke (4 ° C) centrifugation mugihe ukora.

Icyifuzo: Ubushyuhe bwicyumba (15-25 ° C) imikorere, ntuzigere woga urubura hamwe na centrifuge yubushyuhe buke.

2. Kubika icyitegererezo kidakwiye cyangwa ububiko bw'icyitegererezo igihe kirekire.

Icyifuzo: Bika ingero kuri -80 ° C cyangwa uhagarike muri azote yuzuye, kandi wirinde gukoresha inshuro nyinshi;gerageza gukoresha ingero zegeranijwe zivuye muri RNA.

3.Icyitegererezo kidahagije

Icyifuzo: Nyamuneka menya neza ko icyitegererezo nigisubizo cyakazi (Linear Acrylamide) byavanze neza kandi bigashyirwa muminota 10 mubushyuhe bwicyumba (15-25 ° C)

4.Umuhanga wongeyeho nabi

Icyifuzo: Menya neza ko ddH2O yubusa ya RNase yongewe hagati ya membrane yinkingi yo kweza

5.Ubunini budakwiye bwa anhydrous ethanol muri Buffer viRW2

Igitekerezo: Nyamuneka kurikiza amabwiriza, ongeramo ingano yukuri ya anhydrous ethanol kuri Buffer viRW2 hanyuma ubivange neza mbere yo gukoresha ibikoresho。

6.Ikoreshwa ry'icyitegererezo kidakwiye.

Igitekerezo: 200µl yicyitegererezo kuri 500μl ya Buffer viRL.Ingano yikitegererezo irenze izagabanya igipimo cyo gukuramo RNA.

7.Ijwi ridakwiriye cyangwa gusohora kutuzuye.

Igitekerezo: Ingano nziza yinkingi yo kweza ni 30-50μl;niba ingaruka zo gukuraho zidashimishije, birasabwa kongeramo ubushyuhe bwa RNase-Ubusa ddH₂O hanyuma wongere umwanya ushira mubushyuhe bwicyumba, nka 5-10min

8.Inkingi yo kweza ifite ibisigisigi bya Ethanol nyuma yo koza muri Buffer viRW2.

Igitekerezo: Niba Ethanol ikomeje kuguma nyuma yo koza muri Buffer viRW2 hamwe na centrifugation yubusa ya 2min, inkingi yo kweza irashobora gusigara mubushyuhe bwicyumba cya 5min nyuma ya centrifugation yubusa kugirango ikureho Ethanol isigaye.

Kwangirika kwa molekile ya RNA isukuye

Ubwiza bwa RNA yatunganijwe bujyanye nibintu nko kubika icyitegererezo, kwanduza RNase, no gukora.

Isesengura ry'impamvu zisanzwe ：

1.Icyitegererezo cyegeranijwe nticyabitswe mugihe.

Igitekerezo: Niba icyitegererezo kidakoreshejwe mugihe cyo gukusanya, nyamuneka ubike kuri -80 ℃ cyangwa azote yuzuye.Kubikuramo molekile ya RNA, gerageza gukoresha ingero zegeranijwe igihe cyose bishoboka.

2.Ingero zegeranijwe zarakonje kandi zishonga inshuro nyinshi.

Igitekerezo: Irinde gukonjesha no gukonjesha inshuro nyinshi (bitarenze rimwe) mugihe cyo gukusanya no kubika, bitabaye ibyo umusaruro wa acide nucleique uzagabanuka.

3.RNase yatangijwe mucyumba cyo gukoreramo cyangwa nta gants imwe ikoreshwa, masike, nibindi byambarwa.

Igitekerezo: Gukuramo igeragezwa rya molekile ya RNA bikorwa neza mubyumba bitandukanye bya RNA, kandi imbonerahamwe yubushakashatsi isukurwa mbere yubushakashatsi.Wambare uturindantoki hamwe na mask mugihe cyo kugerageza kugirango wirinde kwangirika kwa RNA guterwa na RNase.

4.Ragent yandujwe na RNase mugihe cyo kuyikoresha.

Icyifuzo: Simbuza Virus RNA nshya yo kwigunga kubushakashatsi bujyanye.

5.Indwara ya RNase yanduye ya centrifuge, inama za pipeti, nibindi. Icyifuzo: Menya neza ko imiyoboro ya centrifuge, inama za pipeti, hamwe na pipeti byose ari RNase-Free.

Molekile ya RNA yatunganijwe yagize ingaruka kubushakashatsi bwo hasi

Molekile ya RNA yatunganijwe ninkingi yo kweza izagira ingaruka kubigeragezo byo hepfo niba hari ion nyinshi cyangwa umunyu mwinshi, nka: transcription transcription, Blot y'Amajyaruguru, nibindi .。

1.Hariho ion z'umunyu zisigaye muri molekile ya RNA yatowe.

Icyifuzo: Menya neza ko ingano yukuri ya etanol ya anhydrous yongerewe kuri Buffer viRW2, hanyuma ukarabe inkingi yo kweza kabiri ukurikije umuvuduko ukwiye wa centrifugation kumabwiriza yimikorere ； Niba hakiri ioni yumunyu usigaye, urashobora kongeramo Buffer viRW2 kumurongo wogusukura, hanyuma ukayirekera mubushyuhe bwicyumba cya 5min.Noneho kora centrifugation kugirango ukureho umunyu ion wanduye kurwego runini

2.Hariho Ethanol isigaye muri molekile ya RNA yatowe

Igitekerezo: umaze kwemeza ko inkingi zo kwezwa zogejwe na Buffer viRW2, kora centrifugation yubusa-yubusa ukurikije umuvuduko wa centrifugal kumabwiriza yimikorere.Niba hasigaye etanol isigaye, irashobora gusigara muminota 5 mubushyuhe bwicyumba nyuma ya centrifugation yubusa kugirango ikureho Ethanol isigaye kurwego runini.

Imfashanyigisho:

Igitabo gikubiyemo amabwiriza ya virusi ya RNA