ADN ikuramo virusi / RNA
Iki gikoresho gikwiranye no gukuramo byihuse ADN / RNA ifite isuku nyinshi mu ngero nka nasofaryngeal swabs, swabs ibidukikije, umuco ndengakamere, hamwe na tissue homogenate ndengakamere.Igikoresho gishingiye ku buhanga bwo kweza silika membrane ikuraho ibikenerwa byo gukoresha fenol / chloroform yumuti ukomoka kumyunyu ngugu cyangwa imvura igwa igihe kinini kugirango ikuremo virusi ya ADN / RNA yujuje ubuziranenge.Acide nucleic yabonetse nta mwanda kandi yiteguye gukoreshwa mubigeragezo byo hasi nko kwandukura inyandiko, PCR, RT-PCR, igihe nyacyo PCR, ibihe bizakurikiraho (NGS), hamwe na blot y'Amajyaruguru.
Imiterere yo kubika
Ubike kuri 15 ~ 25 ℃, hanyuma utware ubushyuhe bwicyumba
Ibigize
| Ibigize | 100RXNS |
| Buffer VL | 50 ml |
| Buffer RW | 120 ml |
| RNase idafite ddH2 O. | 6 ml |
| Inkingi yihuse ya RNA | 100 |
| Imiyoboro yo gukusanya (2ml) | 100 |
| Ikusanyirizo rya RNase ridafite ubuntu (1 .5ml) | 100 |
Buffer VL:Tanga ibidukikije bya lysis no guhuza.
Buffer RW:Kuraho poroteyine zisigaye hamwe n’indi myanda.
RNase idafite ddH2O:Kuraho ADN / RNA kuva muri membrane muri spin inkingi.
Inkingi yihuse ya RNA:By'umwihariko adsorb ADN / RNA.
Igikoresho cyo gukusanya 2 ml:Kusanya akayunguruzo.
Ikusanyirizo ridafite RNase 1.5 ml:Kusanya ADN / RNA.
Porogaramu
Nasopharyngeal swabs, ibidukikije, ibidukikije ndengakamere, hamwe na tissue homogenate ndengakamere.
Witegure wenyineials
Inama ya RNette itagira inama, 1.5 ml ya RNase itagira umuyoboro wa centrifuge, centrifuge, mixer ya vortex, na pipettes.
Inzira y'Ubushakashatsi
Kora intambwe zose zikurikira muri kabili ya biosafety.
1 muri make gukusanya imvange hepfo yigituba.
2. Shira Inkingi yihuta ya RNA Inkingi zo gukusanya 2 ml.Hindura imvange kuva ku ntambwe ya 1 kuri FastPure RNA Inkingi, centrifuge kuri 12,000 rpm (13,400 × g) muminota 1, hanyuma ujugunye filtrate.
3. Ongeramo 600 μl ya Buffer RW kuri FastPure RNA Inkingi, centrifuge kuri 12,000 rpm (13,400 × g) kumasegonda 30, hanyuma ujugunye filtrate.
4. Subiramo Intambwe ya 3.
5. Centrifuge inkingi yubusa kuri 12,000 rpm (13.400 × g) kuminota 2.
6. Witonze wimure Inkingi yihuta ya RNA mumashanyarazi mashya ya RNase idafite miriyoni 1.5 (yatanzwe mubikoresho), hanyuma ongeramo 30 - 50 μl ya RNase idafite ddH2O hagati ya membrane udakora ku nkingi.Emera guhagarara mubushyuhe bwicyumba kuminota 1 na centrifuge kuri 12,000 rpm (13,400 × g) kuminota 1.
7. Hagarika Inkingi Yihuta ya RNA.ADN / RNA irashobora gukoreshwa muburyo butaziguye mubisubizo byakurikiyeho, cyangwa ikabikwa kuri -30 ~ -15 ° C mugihe gito cyangwa -85 ~ -65 ° C mugihe kirekire.
Inyandiko
Kubushakashatsi ukoreshe gusa.Ntabwo ari ugukoresha muburyo bwo gusuzuma.
1. Kuringaniza ingero n'ubushyuhe bw'icyumba mbere.
2. Virusi zirandura cyane.Nyamuneka menya neza ko ingamba zose z'umutekano zafashwe mbere yo kugerageza.
3. Irinde gukonjesha no gukonjesha icyitegererezo, kuko ibyo bishobora gutera kwangirika cyangwa kugabanya umusaruro wa ADN / RNA yakuweho virusi.
4. Ibikoresho byateguwe ubwabyo birimo inama ya RNase itagira inama, 1.5 mili ya RNase idafite imiyoboro ya centrifuge, centrifuge, mixer ya vortex, na pipettes.
5. Mugihe ukoresheje ibikoresho, ambara ikote rya laboratoire, uturindantoki twa latx ikoreshwa, hamwe na mask ikoreshwa kandi ukoreshe ibikoresho bidafite RNase kugirango ugabanye ibyago byo kwanduza RNase.
6. Kora intambwe zose mubushyuhe bwicyumba keretse byateganijwe ukundi.
Urwego rukoreshwa
