Kiti ya Magnetic Universal Viral DNA/RNA HYD521
Kiti ya Magnetic Universal Viral DNA/RNA ndi yoyenera kutulutsa asidi wa viral nucleic acid m'magazi athunthu kapena m'madzi amthupi opanda maselo (monga seramu, plasma, madzi a m'mitsempha, swabs za m'mphuno/pharyngeal, madzi osambira a alveolar, ndi zina zotero). Njirayi imagwiritsa ntchito ukadaulo woyeretsa mikanda ya magnetic bead, ndipo palibe chochotsa cha phenol chloroform chomwe chikufunika panthawi yochotsa. Ndi yotetezeka, yopanda poizoni komanso yachangu. Chogulitsa chomwe chapezeka chingagwiritsidwe ntchito mwachindunji mu PCR, qPCR, sequencing ya m'badwo wachiwiri ndi zoyeserera zina. Mukagwiritsidwa ntchito ndi zida zochotsera za mikanda ya magnetic, kutulutsa ma nucleic acids ambiri kumatha kuchitika.
Zambiri za malonda
| Zigawo | 48T |
| Lysis ndi Binding Solution | 33 mL |
| Kuyimitsidwa kwa mkanda wamaginito | 1.1 mL |
| Chotsukira chotsukira A | 40 mL |
| Chotsukira chotsukira B | 80 mL |
| Yankho la Elution | 10 mL |
| Mapuloteni K | 1.1 mL |
Mikhalidwe Yosungira
Proteinase K iyenera kusungidwa pa kutentha kwa 2 ~ 8 ℃ ndipo zosakaniza zina ziyenera kusungidwa kutentha kwa chipinda. Nthawi yosungiramo zinthu ndi miyezi 18.
Kufotokozera kwa Chitsanzo
1. Mitundu ya zitsanzo zomwe zimagwiritsidwa ntchito: magazi athunthu, seramu, plasma, madzi a m'mitsempha ya ubongo, swabs za m'mphuno/m'mero, madzi osambira a m'matumbo ndi zitsanzo zina.
2. Kusunga ndi kunyamula zitsanzo: Zitsanzo zitha kugwiritsidwa ntchito poyesa nthawi yomweyo kapena kusungidwa pa -70℃ kapena kutsika kuti ziyesedwe. Nthawi yosungira ndi miyezi 6. Kuzizira mobwerezabwereza ndi kusungunuka kuyenera kupewedwa. Zitsanzo zimanyamulidwa ndi unyolo wozizira.
3. Zofunikira pa kuzizira ndi kusungunuka: Kuzizira ndi kusungunuka mwachangu, pewani kuzizira ndi kusungunuka mobwerezabwereza.
Ntchito
Musanayesere, yang'anani ngati yankholo lili ndi mvula komanso ngati mikanda ya maginito ikhoza kuyimitsidwanso.
1. Tumizani 200~300 μL ya chitsanzo ku chubu cha centrifuge cha 1.5 mL, onjezerani 600 μL ya Lysis and Binding Solution, 20 μL ya Proteinase K, ndi 20 μL ya magnetic bead suspension motsatizana, ndi vortex pa liwiro lapamwamba kwa masekondi 30. Ikani pa 50℃ kwa mphindi 5~7. Ngati thermostat ilibe ntchito yosinthasintha, ikani vortex katatu panthawi yopumira, nthawi iliyonse kwa masekondi 15.
2. Ikani mu 1.5 mL magnetic stand kuti mulekanitse maginito mpaka yankho likhale loyera komanso lowonekera, ndipo taya yankholo.
3. Onjezani 700 μL ya wash buffer A, vortex kwa masekondi 10 kuti muswe mikanda ya maginito, kenako muyike ku malo oimika maginito kuti mulekanitse maginito mpaka yankho litayera, ndipo taya yankholo.
4. Onjezani 700 μL ya chotsukira madzi B, vortex kwa masekondi 10 kuti muswe mikanda ya maginito, sinthani ku malo oimika maginito kuti mulekanitse maginito mpaka yankho litayera, ndikutaya yankholo.
5. Onjezani 700 μL ya chotsukira cha B kachiwiri, pukutani vortex kwa masekondi 10 kuti muswe mikanda ya maginito, sungani ku malo oimikapo maginito kuti mulekanitse maginito mpaka yankho litayera, ndikutaya yankholo.
6. Thirani mpweya pang'ono kuti musonkhanitse madontho pakhoma la chubu, muwasamutse ku choyimilira cha maginito kuti chiwonekere bwino, ndikuyamwa madzi otsala. Umitsani mpweya kwa mphindi 3-5.
Dziwani: Zotsalira za ethanol zimaletsa zochita za enzyme pambuyo pake, choncho onetsetsani kuti ethanol yatha kwathunthu ikauma. Musaume kwa nthawi yayitali kuti musawononge zotsatira za elution.
7. Onjezani 40~100 μL ya yankho la Elution, vortex pa liwiro lalikulu kwa mphindi 2~3 kuti muswe mikanda ya maginito. Ikani pa kutentha kwa 60℃ kwa mphindi 5, kenako vortex pa liwiro lalikulu kwa masekondi 60.
8. Ikani centrifuge nthawi yomweyo kuti mutenge madziwo kuchokera pachivundikiro cha chubu ndikusamutsa ku choyikapo maginito mpaka mikanda ya maginito italowa bwino. Kenako sungani madziwo mosamala ku chubu chatsopano cha centrifuge kuti mupeze yankho la nucleic acid.
9. Yankho la nucleic acid likhoza kusungidwa pa -20°C kuti lisungidwe kwa kanthawi kochepa ndi -80°C kuti lisungidwe kwa nthawi yayitali.
Zolemba
1. Ma buffer osiyanasiyana omwe ali mu kit iyi ali ndi mchere wa guanidine. Kuti mukhale otetezeka komanso athanzi, chonde valani jekete la labu ndi magolovesi otayidwa. Ndipo gwiritsani ntchito motsatira njira zodzitetezera. Musalole kuti buffer ikhudze khungu, maso ndi nembanemba ya mucous. Ngati zitachitika, chonde sambani nthawi yomweyo ndi madzi ambiri ndikupita kuchipatala.
2. Ngati yankho lafika pachimake, liyenera kutsukidwa m'madzi otentha 30°C mpaka phulusa litasungunuka kwathunthu musanagwiritse ntchito.
3. Ngati chotsukira cha maginito chazizira, musachigwiritse ntchito.
4. Ma buffer osiyanasiyana omwe ali mu kit iyi ali ndi mchere wa guanidine. Musagwiritse ntchito mankhwala ophera tizilombo toyambitsa matenda monga sodium hypochlorite kuti muwachiritse, apo ayi mpweya woipa udzatulutsidwa ndipo uyenera kuonedwa ngati zinyalala zachipatala.
5. Pakhoza kukhala mikanda yotsalira ya maginito panthawi yopumira. Yesetsani kupewa kupuma mikanda ya maginito mukatulutsa madzi.