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Akagari Igiteranyo Cyuzuye RNA Igikoresho Cyuzuye RNA Yigunga Ibikoresho byo muri selire

Ibisobanuro:

RNA isukuye cyane kandi yujuje ubuziranenge RNA irashobora kuboneka mumasoko atandukanye yimico muminota 11.

RNase-Ubuntu

Bikorera mubushyuhe bwicyumba (15-25 ℃)

Hamwe na ADN-Isukura Inkingi

Umusaruro mwinshi wa RNA

Byihuse: Kurangiza gukuramo muminota 11

Umutekano: Nta miti kama

imbaraga


  • :
  • Ibicuruzwa birambuye

    Ibicuruzwa

    Ibibazo

    Ibisobanuro

    Iki gikoresho gikoresha inkingi ya spin hamwe na formula yakozwe na Foregene, ishobora gukuramo neza-isuku nini kandi nziza-yuzuye ya RNA ivuye mu ngirabuzimafatizo zifite 96, 24, 12, na 6-nziza.

    Igikoresho gitanga ADN isukura neza, ishobora gutandukanya byoroshye na lysate ndengakamere, guhuza no gukuraho ADN genomic.Igikorwa kiroroshye kandi gitwara igihe.

    Twe RNA-Inkingi yonyine irashobora guhuza neza RNA na formula idasanzwe.Umubare munini wintangarugero urashobora gutunganyirizwa icyarimwe.

    Ibigize ibikoresho

    Ibigize ibikoresho RE-03111 RE-03114
    50 T. 200 T.
    Buffer cRL1 * 25 mL 100 mL
    Buffer cRL2 15 mL 60 mL
    Buffer RW1 * 25 mL 100 mL
    Buffer RW2 24 mL 96 mL
    RNase Yubusa ddH2O 10 mL 40 mL
    RNA-Inkingi gusa 50 200
    Inkingi ya ADN 50 200
    Amabwiriza 1 1

    * Nyamuneka wambare uturindantoki kandi ufate ingamba zo kubarinda mugihe cyo kubaga kuko Buffer cRL1 na Buffer RW1 zirimo umunyu wa chaotropique.

    Ibiranga ibyiza

    Process Ibikorwa byose bikorerwa mubushyuhe bwicyumba (15-25 ℃), nta bwogero bwa barafu hamwe na centrifugation yubushyuhe buke.
    Kit Ibikoresho byose ni RNase-Ubuntu, nta mpamvu yo guhangayikishwa no kwangirika kwa RNA.
    Inkingi ya ADN-Isukura ihuza ADN cyane cyane, kugirango igikoresho gishobora gukuraho ADN yanduye utiriwe wongeraho DNase.
    Umusaruro mwinshi wa RNA: RNA-Inkingi gusa hamwe na formula idasanzwe irashobora kweza neza RNA.
    Speed ​​Umuvuduko wihuse: byoroshye gukora kandi birashobora kurangira muminota 11.
    Umutekano: Nta reagent ikenewe.
    Quality Ubwiza buhanitse: RNA isukuye ifite isuku nyinshi, idafite poroteyine n’indi mwanda, kandi irashobora guhura nubushakashatsi butandukanye bwakurikiyeho.

    ibyiza bya foregene RNA Isolation kit

    Porogaramu

    Irakwiriye gukuramo no kweza RNA yose muri selile zifite imico muri plaque 96, 24, 12, na 6.

    Urujya n'uruza rw'akazi

    selile yose RNA

    Igishushanyo

    Akagari Igiteranyo Cyuzuye RNA Igikoresho Cyakazi Flow1

    Igishushanyo cya batiri ya agarose gel ya Cell Total RNA Isolation Kit yavuzeko imibare itandukanye ya selile yavuzwe haruguru, gukuramo 20μl, gufata 2μl isukuye yose hamwe RNA 1%.

    Ububiko nubuzima bwiza

    Igikoresho gishobora kubikwa amezi 12 mubushyuhe bwicyumba (15-25 ℃) cyangwa 2-8 ℃ mugihe kinini (amezi 24).

    Buffer cRL1 irashobora kubikwa kuri 4 ℃ ukwezi 1 nyuma yo kongeramo 2-hydroxy-1-Ethanethiol (bidashoboka).


  • Mbere:
  • Ibikurikira:

  • RNA ntabwo yakuweho cyangwa umusaruro wa RNA ni muke

    Hariho ibintu byinshi bigira ingaruka kumikorere yo gukira, nka: tissue sample ya RNA ibirimo, uburyo bwo gukora, ingano ya elution, nibindi.

    1. Kwiyuhagira urubura cyangwa cryogenic (4 ° C) centrifugation yakozwe mugihe cyo gukora.

    Icyifuzo: Kora ku bushyuhe bwicyumba (15-25 ° C) mugihe cyose, ntukarabe hamwe na centrifuge mubushyuhe buke.

    2. Kubika icyitegererezo kidakwiye cyangwa igihe cyo kubika urugero.

    Icyifuzo: Bika ingero kuri -80 ° C cyangwa uhagarike muri azote yuzuye kandi wirinde gukoresha inshuro nyinshi;gerageza gukoresha tissue nshya cyangwa selile zifite imico yo gukuramo RNA.

    3. Lisisi ntangarugero idahagije.

    Icyifuzo: Mugihe ingirabuzima fatizo, menya neza ko ingirangingo zahujwe bihagije kandi ko ingirabuzimafatizo zigabanyijemo bihagije kugirango zisobanure irekurwa rya RNA.

    4. Umuhanga ntabwo yongeweho neza.

    Icyifuzo: Emeza ko ddH idafite RNase2O yongeweho ibitonyanga hagati yo kweza inkingi ya membrane.

    5. Ingano yukuri ya Ethanol yuzuye ntabwo yongewe kuri Buffer RL2 cyangwa Buffer RW2.

    Icyifuzo: Kurikiza amabwiriza, ongeramo ingano yukuri ya Ethanol yuzuye kuri Buffer RL2 na Buffer RW2 hanyuma uvange neza mbere yo gukoresha ibikoresho.

    6. Dissue sample dosage ntabwo ikwiye.

    Icyifuzo: Koresha 10-20 mg ya tissue cyangwa (1-5) × 106selile kuri 500 μl buffer RL1, kuko gukoresha tissue birenze urugero bishobora kugabanya gukuramo RNA.

    7. Ingano yo gukuraho idakwiye cyangwa gusiba kutuzuye.

    Icyifuzo: Ingano yo gukuraho inkingi yo kweza ni 50-200 μl;niba ingaruka zo gukuraho zidashimishije, birasabwa kongera igihe cyo gushyira ubushyuhe bwicyumba nyuma yo kongeramo ubushyuhe bwa RNase-Ubusa ddH2O, urugero kuminota 5-10.

    8.Inkingi yo kweza ifite ibisigisigi bya Ethanol nyuma yo gukaraba Buffer RW2.

    Icyifuzo: Niba hari ibisigisigi bya Ethanol nyuma yo gukaraba Buffer RW2, centrifugation yubusa ya 1min, igihe cyo gukora igituba cyubusa centrifugation gishobora kwiyongera kugeza 2min, cyangwa inkingi yo kweza irashobora gushyirwa mubushyuhe bwicyumba kuminota 5 kugirango ikureho Ethanol isigaye.

    RNA isukuye irateshwa agaciro

    Ubwiza bwa RNA yatunganijwe bujyanye nibintu nko kubungabunga icyitegererezo, kwanduza RNase, na manipulation, nibindi.

    1. Ingero z'imyenda ntizibikwa mugihe.

    Icyifuzo: Niba ingero za tissue cyangwa selile zidakoreshejwe mugihe gikwiye nyuma yo kwegeranya, hita ubika neza kuri -80 ° C cyangwa azote yuzuye.Gukuramo RNA, koresha tissue nshya yafashwe cyangwa selile selile igihe cyose bishoboka.

    2. Gusubiramo inshuro nyinshi gukonjesha ingero za tissue.

    Icyifuzo: Mugihe ubitse ingero za tissue, nibyiza ko ubicamo uduce duto kugirango ubungabunge, hanyuma ukuremo kimwe mubice mugihe ubikoresheje kugirango wirinde gukonjesha inshuro nyinshi no kwangirika kwa RNA.

    3. RNase itangizwa cyangwa itambaye uturindantoki twajugunywe, masike, nibindi mugihe cyo gukora.

    Icyifuzo: Ubushakashatsi bwo gukuramo RNA bukorwa neza mubyumba bitandukanye bya manipulation ya RNA kandi ameza arasukurwa mbere yubushakashatsi.

    Wambare uturindantoki hamwe na masike mugihe cyo kugerageza kugirango ugabanye kwangirika kwa RNA guterwa no kwinjiza RNase.

    4. Reagents zandujwe na RNase mugihe cyo kuyikoresha.

    Icyifuzo: Simbuza inyamaswa nshya Igiteranyo cya RNA cyo kwigunga kubushakashatsi bujyanye.

    5. Imiyoboro ya centrifuge, inama, nibindi bikoreshwa muri manipulation ya RNA byanduye na RNase.

    Icyifuzo: Emeza ko imiyoboro ya centrifuge, inama, imiyoboro, nibindi bikoreshwa mugukuramo RNA byose ni RNase-Ubuntu.

    RNA yatunganijwe neza igira ingaruka kubushakashatsi bwo hasi

    RNA yasukuwe ninkingi yo kweza, niba ion yumunyu, proteyine nini nini cyane bizagira ingaruka kubigeragezo byo hasi, nka: transcription transcription, Blot y'Amajyaruguru nibindi.

    1. RNA yakuweho ifite ibisigazwa byumunyu.

    Icyifuzo: Emeza ko ingano yukuri ya Ethanol yongewe kuri Buffer RW2 hanyuma ukore inkingi 2 yoza inkingi kumuvuduko wa centrifugal yerekanwe kubikorwa;niba hari ibisigazwa byumunyu ion, usige inkingi yo kweza kuri Buffer RW2 muminota 5 mubushyuhe bwicyumba hanyuma ukore centrifugation kugirango ukureho umwanda mwinshi.

    2. Ibisigisigi bya Ethanol muri RNA yatowe.

    Icyifuzo: Emeza ko nyuma yo gukaraba buffer RW2, kora ibikorwa byubusa bya centrifugation yubusa kumuvuduko wa centrifugation yerekanwe kubikorwa, ongera igihe cyibikorwa bya centrifugation yubusa kugeza min 2 niba hakiri ibisigisigi bya Ethanol, cyangwa ubireke mubushyuhe bwicyumba kuri m 5

    Andika ubutumwa bwawe hano hanyuma utwohereze