Ikhithi yeMagnetic Universal Viral DNA/RNA HYD521
I-Magnetic Universal Viral DNA/RNA Kit ifanelekile ekukhupheni i-viral nucleic acid egazini elipheleleyo okanye kulwelo lomzimba olungenaseli (ezifana ne-serum, i-plasma, ulwelo lwe-cerebrospinal, i-nasal/pharyngeal swabs, ulwelo lokuhlamba i-alveolar, njl.njl.). Le ndlela isebenzisa iteknoloji yokucoca ii-magnetic bead, kwaye akukho mfuneko yokukhupha i-phenol chloroform enetyhefu ngexesha lenkqubo yokukhupha. Ikhuselekile, ayinatyhefu kwaye iyakhawuleza. Imveliso efunyenweyo ingasetyenziswa ngokuthe ngqo kwi-PCR, qPCR, ulandelelwano lwesizukulwana sesibini kunye nezinye iimvavanyo. Xa isetyenziswa nezixhobo zokukhupha ii-magnetic bead ezenzekelayo, ukukhupha ii-nucleic acids eziphezulu kunokufezekiswa.
Ulwazi lwemveliso
| Izinto eziyinxalenye | 48T |
| Isisombululo seLysis kunye neBinding | 33 ml |
| Ukumiswa kwe-magnetic bead | 1.1 mL |
| Isithinteli sokuhlamba impahla A | 40 ml |
| Isithinteli sokuhlamba ukuhlamba B | 80 mL |
| Isisombululo se-Elution | 10 ml |
| Iproteni K | 1.1 mL |
Iimeko Zokugcina
I-Proteinase K kufuneka igcinwe kumaqondo obushushu angama-2 ~ 8℃ kwaye ezinye izithako mazigcinwe kumaqondo obushushu egumbi. Ixesha lokugcinwa kweproteni ziinyanga ezili-18.
Inkcazo yeSampuli
1. Iindidi zeesampuli ezisebenzayo: igazi elipheleleyo, i-serum, i-plasma, ulwelo lwe-cerebrospinal, ii-nasal/pharyngeal swabs, ulwelo lokuhlamba i-alveolar kunye nezinye iisampuli.
2. Ukugcinwa kunye nokuthuthwa kweesampuli: Iisampuli zingasetyenziselwa ukuvavanywa ngoko nangoko okanye zigcinwe kwi -70℃ okanye ngaphantsi ukuze kuvavanywe. Ixesha lokugcina ziinyanga ezi-6. Ukuqandisa okuphindaphindiweyo kunye nokunyibilika kufuneka kuthintelwe. Iisampuli zithuthwa ngetsheyini ebandayo.
3. Iimfuneko zokuqandisa nokunyibilikisa: Ukuqandisa nokunyibilikisa ngokukhawuleza, kuphephe ukuqandisa nokunyibilikisa ngokuphindaphindiweyo.
Umsebenzi
Ngaphambi kovavanyo, jonga ukuba isisombululo sinayo na imvula kwaye ingaba iintsimbi zemagnethi zinokuphinda zixhonywe.
1. Dlulisa i-200~300 μL yesampuli kwityhubhu ye-centrifuge ye-1.5 mL, yongeza i-600 μL yeLysis and Binding Solution, i-20 μL yeProteinase K, kunye ne-20 μL ye-magnetic bead suspension ngokulandelelana, kunye ne-vortex ngesantya esiphezulu imizuzwana engama-30. Fukama kwi-50℃ imizuzu emi-5~7. Ukuba i-thermostat ayinawo umsebenzi we-oscillation, fudumeza i-vortex izihlandlo ezi-3 ngexesha le-incubation, ixesha ngalinye imizuzwana eli-15.
2. Dlulisa kwi-1.5 mL magnetic stand ukuze ufumane ulwahlulo lwe-magnetic de isisombululo sibe sicacile kwaye sibonakala, uze usilahle isisombululo.
3. Yongeza i-700 μL ye-wash buffer A, i-vortex imizuzwana eli-10 ukuze uqhekeze ii-magnetic beads, uze uyithumele kwindawo yokuma ye-magnetic ukuze uhlukanise i-magnetic de isisombululo sibe mhlophe, uze ulahle isisombululo.
4. Yongeza i-700 μL ye-wash buffer B, i-vortex imizuzwana eli-10 ukuze uqhekeze ii-magnetic beads, uyidlulisele kwi-magnetic stand ukuze uhlukanise i-magnetic de isisombululo sibe mhlophe, uze ulahle isisombululo.
5. Yongeza i-700 μL ye-wash buffer B kwakhona, faka i-vortex imizuzwana eli-10 ukuze uqhekeze ii-magnetic beads, udlulisele kwi-magnetic stand ukuze uhlukanise i-magnetic de isisombululo sibe mhlophe, uze ulahle isisombululo.
6. Yifake kwi-centrifuge okwethutyana ukuze uqokelele amathontsi eludongeni lwetyhubhu, uyidlulisele kwindawo enomtsalane ukuze icace, kwaye uyifunxe intsalela yolwelo. Yomisa ngomoya imizuzu emi-3-5.
Qaphela: Intsalela ye-ethanol iya kuthintela ukusabela kwe-enzyme okulandelayo, ngoko ke qiniseka ukuba i-ethanol iyaphela ngokupheleleyo xa yomile. Musa ukomisa ixesha elide ukuze ungachaphazeli isiphumo sokukhupha i-ethanol emva koko.
7. Yongeza i-40~100 μL yesisombululo se-Elution, i-vortex ngesantya esiphezulu imizuzu emi-2~3 ukuze uqhekeze iintsimbi zemagnethi. Yifukamele kwi-60℃ imizuzu emi-5, uze uyifukamele ngesantya esiphezulu imizuzwana engama-60.
8. Faka i-centrifuge kwangoko ukuze uqokelele ulwelo oluvela kwisigqubuthelo setyhubhu uze uludlulisele kwi-magnetic rack de ii-magnetic beads zifunxwe ngokupheleleyo. Emva koko dlulisela ulwelo ngononophelo kwityhubhu entsha ye-centrifuge ukuze ufumane isisombululo se-nucleic acid.
9. Isisombululo se-nucleic acid singagcinwa kwi--20°C ukuze sigcinwe ixesha elifutshane kunye ne--80°C ukuze sigcinwe ixesha elide.
Amanqaku
1. Ii-buffers ezahlukeneyo kule khithi ziqulethe i-guanidine salts. Ukuze ukhuseleke kwaye ube sempilweni, nxibe ijazi lelebhu kunye neeglavu ezilahlwayo. Kwaye uphathe ngokwemigaqo yokhuseleko. Musa ukuvumela i-buffer idibane nolusu, amehlo kunye ne-mucous membranes. Ukuba kuyenzeka, nceda uhlambe ngokukhawuleza ngamanzi amaninzi kwaye ufumane uncedo lwezonyango.
2. Ukuba isisombululo siyavuthuluka, kufuneka sihlanjwe ngamanzi angama-30°C de i-precipitate inyibilike ngokupheleleyo ngaphambi kokuba sisetyenziswe.
3. Ukuba i-magnetic bead suspension ikhenkcezisiwe, ungayisebenzisi.
4. Ii-buffers ezahlukeneyo kule khithi ziqulethe iityuwa ze-guanidine. Musa ukusebenzisa izibulali-ntsholongwane ezikhupha ioksijini ezifana ne-sodium hypochlorite ukuzinyanga, kungenjalo iigesi ezinobuthi ziya kukhutshwa kwaye kufuneka ziphathwe njengenkunkuma yezonyango.
5. Kusenokubakho iintsimbi zemagnethi eziseleyo ngexesha lokucoca. Zama ukuphepha ukuphefumla iintsimbi zemagnethi xa ufutha iisampulu.