Qalabka Soo saarista DNA/RNA Viral
Qalabkani wuxuu ku habboon yahay in si degdeg ah looga soo saaro DNA/RNA fayraska nadiifka ah ee shaybaarada sida suufka naasopharyngeal, swabs deegaanka, kor u kaca dhaqanka unugyada, iyo unugyo isku-dhafan oo sarreeya.Qalabku wuxuu ku salaysan yahay tignoolajiyada nadiifinta xuubka silica taasoo meesha ka saaraysa baahida loo qabo isticmaalka dareerayaasha phenol/chloroform organic ama roobabka alkohol waqti-qaada si loo soo saaro fayraska DNA/RNA oo tayo sare leh.Asiidhyada nucleic-ka ah ee la helay waa kuwo ka nadiif ah wasakh waxayna diyaar u yihiin in loo isticmaalo tijaabooyinka hoose sida qoraalka dib u dhigista, PCR, RT-PCR, PCR-waqtiga-dhabta ah, isku xigxiga jiilka xiga (NGS), iyo Northern blot.
Xaaladaha kaydinta
Ku kaydi at 15 ~ 25 ℃, oo ku qaado heerkulka qolka
Qaybaha
| Qaybaha | 100RXNS |
| Buffer VL | 50 ml |
| Buffer RW | 120 ml |
| RNase-free ddH2 O | 6 ml |
| FastPure RNA Columns | 100 |
| Tuubooyinka Aruurinta (2ml) | 100 |
| Tubooyinka Ururinta ee bilaa RNase (1.5ml) | 100 |
Buffer VL:Bixi jawi loogu talagalay lysis iyo xidhitaanka.
Bakhaar RW:Ka saar borotiinnada haraaga ah iyo wasakhda kale.
RNase-free ddH2O:Elute DNA/RNA oo ka soo baxa xuubka laf dhabarta.
Tiirarka RNA FastPure:Gaar ahaan adsorb DNA/RNA.
Tuubooyinka Aruurinta 2 ml:Ururi sifaynta
Tubooyinka Ururinta ee bilaa RNase 1.5 ml:Ururi DNA/RNA.
Codsiyada
Naasopharyngeal swabs, swabs deegaanka, supernatants dhaqanka unug, iyo unug-homogenate supernatants.
Mater is-diyaarsadayilbaxnimo
Talooyin pipette-free RNase, 1.5 ml RNase-free tuubooyinka centrifuge, centrifuge, qase vortex, iyo pipettes.
Habka tijaabada
Ku samee dhammaan tillaabooyinka soo socda gudaha golaha badbaadada noolaha.
1. Ku dar 200 μl ee muunada tuubada sentifuge-free RNase (ka samaysan PBS ama 0.9% NaCl haddii ay dhacdo muunad ku filnayn), ku dar 500 μl ee Buffer VL, si fiican u walaaq adigoo ku wareegaya 15 - 30 sek, iyo centrifuge si kooban si aad u ururiso isku dhafka hoose ee tuubada.
2. Ku rid FastPure RNA Columns gudaha Tubooyinka Aruurinta 2 ml.Ku wareeji isku dhafka Talaabada 1 una wareeji FastPure RNA Columns, centrifuge at 12,000 rpm (13,400 × g) 1 daqiiqo, oo tuur shaandhada.
3. Ku dar 600 μl ee Buffer RW FastPure RNA Columns, centrifuge at 12,000 rpm (13,400 × g) 30 sekan, oo tuur shaandhada.
4. Ku celi talaabada 3aad.
5. Ka dhig tiirka madhan 12,000 rpm (13,400 × g) ilaa 2 daqiiqo.
6. Si taxadir leh ugu wareeji FastPure RNA Columns 1.5 ml Tubooyinka Ururinta bilaa RNase ee cusub, oo ku dar 30 – 50 μl oo ddH2O ah oo RNase-la'aan ah badhtanka xuubka adigoon taaban tiirka.U oggolow inaad ku istaagto heerkulka qolka 1 daqiiqo iyo sentrifuge at 12,000 rpm (13,400 × g) 1 daqiiqo.
7. Tuur FastPure RNA Columns.DNA/RNA waxaa si toos ah loogu isticmaali karaa baaritaanada xiga, ama lagu kaydin karaa -30 ~ -15°C mudo gaaban ama -85 ~-65°C mudo dheer.
Xusuusin
Isticmaalka cilmi-baarista kaliya.Looma isticmaalo hababka ogaanshaha.
1. Horay u dhig shaybaarada heerkulka qolka.
2. Fayrasyadu aad bay u faafaan.Fadlan hubi in dhammaan taxaddarrada badbaadada lagama maarmaanka ah la qaaday tijaabada ka hor.
3. Iska ilaali qaboojinta soo noqnoqda iyo dhalaalidda shaybaarka, maxaa yeelay tani waxay u horseedi kartaa hoos u dhac ama hoos u dhaca wax-soosaarka fayraska la soo saaray ee DNA/RNA.
4. Qalabka is-diyaariyay waxaa ka mid ah talooyinka pipette-free RNase, 1.5 ml tuubooyinka sentrifuge-free RNase, centrifuge, qase vortex, iyo pipettes.
5. Markaad isticmaalayso xirmada, xidho jaakad shaybaadhka, galoofyada cinjirka ah ee la tuuri karo, iyo maaskaro la tuuri karo oo isticmaal waxyaallaha RNase-ka la'aanta ah si loo yareeyo khatarta wasakheynta RNase.
6. Ku samee dhammaan tillaabooyinka heerkulka qolka ilaa si kale loo cayimo.
Farsamada & Socodka Shaqada
