Viral DNA/RNA Extraction Kit
Chidachi ndi choyenera kutulutsa mwachangu kwa DNA/RNA ya virus ya DNA/RNA kuchokera ku zitsanzo monga ma swabs a nasopharyngeal, swabs zachilengedwe, supernatants cell culture, ndi minofu homogenate supernatants.Zidazi zimachokera ku teknoloji yoyeretsa nembanemba ya silika yomwe imathetsa kufunika kogwiritsa ntchito phenol/chloroform organic solvents kapena mvula yamkuntho yomwe imatenga nthawi kuti ichotse ma virus a DNA/RNA apamwamba kwambiri.Ma nucleic acid omwe amapezeka amakhala opanda zodetsa ndipo ali okonzeka kugwiritsidwa ntchito poyesa kutsika kwapansi monga kulembera kumbuyo, PCR, RT-PCR, PCR yeniyeni, kutsatizana kwa mibadwo yotsatira (NGS), ndi Northern blot.
Zosungirako
Sungani pa 15 ~ 25 ℃, ndikuyendetsa kutentha
Zigawo
| Zigawo | Mtengo wa 100RXNS |
| Mtengo VL | 50 ml |
| Mtengo RW | 120 ml |
| RNase-free ddH2 O | 6 ml pa |
| FastPure RNA Columns | 100 |
| Machubu Osonkhanitsira (2ml) | 100 |
| Machubu Osonkhanitsira Aulere a RNase(1 .5ml) | 100 |
Bwalo la VL:Perekani chilengedwe cha lysis ndi kumanga.
Mtengo RW:Chotsani mapuloteni otsalira ndi zonyansa zina.
RNase-free ddH2O:Elute DNA/RNA kuchokera pa nembanemba pagawo lozungulira.
Mizati ya FastPure RNA:Makamaka adsorb DNA/RNA.
Machubu osonkhanitsira 2 ml:Sungani zosefera.
Machubu Osonkhanitsira Aulere a RNase 1.5 ml:Sungani DNA/RNA.
Mapulogalamu
Nasopharyngeal swabs, swabs zachilengedwe, supernatants cell culture, ndi minofu homogenate supernatants.
Wodzikonzekera Materizi
Malangizo a mapaipi opanda RNase, 1.5 ml RNase-free centrifuge chubu, centrifuge, vortex mixer, ndi pipettes.
Njira Yoyesera
Chitani zonse zotsatirazi mu kabati ya biosafety.
1. Onjezani 200 μl yachitsanzo ku chubu chopanda RNase cha centrifuge (pangani ndi PBS kapena 0.9% NaCl ngati sichikukwanira) onjezerani 500 μl ya Buffer VL, sakanizani bwino ndi vortexing kwa 15 - 30 sec, ndi centrifuge mwachidule kusonkhanitsa osakaniza pansi pa chubu.
2. Ikani Mizati ya FastPure RNA mu Machubu Osonkhanitsa 2 ml.Tumizani osakaniza kuchokera ku Gawo 1 kupita ku FastPure RNA Columns, centrifuge pa 12,000 rpm (13,400 × g) kwa 1 min, ndikutaya filtrate.
3. Onjezani 600 μl ya Buffer RW ku FastPure RNA Columns, centrifuge pa 12,000 rpm (13,400 × g) kwa masekondi 30, ndikutaya kusefa.
4. Bwerezani Gawo 3.
5. Centrifuge ndime yopanda kanthu pa 12,000 rpm (13,400 × g) kwa 2 min.
6. Samutsani mosamala Magawo a FastPure RNA mu Machubu Osonkhanitsira Opanda RNase atsopano 1.5 ml (operekedwa mu kit), ndi kuwonjezera 30 - 50 μl ya RNase-free ddH2O pakati pa nembanemba popanda kukhudza gawolo.Lolani kuyimirira kutentha kwa 1 min ndi centrifuge pa 12,000 rpm (13,400 × g) kwa 1 min.
7. Tayani Zida za FastPure RNA.DNA / RNA ikhoza kugwiritsidwa ntchito mwachindunji pazoyesa zotsatila, kapena kusungidwa pa -30 ~ -15 ° C kwa nthawi yochepa kapena -85 ~ -65 ° C kwa nthawi yaitali.
Zolemba
Kugwiritsa ntchito kafukufuku kokha.Osagwiritsidwa ntchito pozindikira matenda.
1. Lumikizani zitsanzo ku kutentha kwa chipinda pasadakhale.
2. Ma virus ndi opatsirana kwambiri.Chonde onetsetsani kuti njira zonse zodzitetezera zikutsatiridwa musanayese.
3. Pewani kuzizira mobwerezabwereza ndi kusungunuka kwa chitsanzo, chifukwa izi zingayambitse kuwonongeka kapena kuchepetsa zokolola za DNA / RNA yotulutsidwa.
4. Zida zodzikonzera zokha zimaphatikizapo malangizo a pipette a RNase, 1.5 ml RNase-free centrifuge tubes, centrifuge, vortex mixer, ndi pipettes.
5. Mukamagwiritsa ntchito zidazi, valani chovala cha labu, magolovesi a latex otayidwa, ndi chigoba chotayira ndipo gwiritsani ntchito zinthu zopanda RNase kuti muchepetse kuopsa kwa RNase.
6. Chitani masitepe onse kutentha kwa chipinda pokhapokha ngati tafotokozera.
Mechanism & ntchito
