Ngwa DNA/RNA Magnetik zuru ụwa ọnụ nke nje HYD521
Ngwa DNA/RNA nke Magnetic Universal Viral dị mma maka iwepụ nucleic acid nje site na ọbara dum ma ọ bụ mmiri ahụ na-enweghị sel (dịka serum, plasma, mmiri cerebrospinal, swabs imi/pharyngeal, mmiri ịsa ahụ alveolar, wdg). Usoro a na-eji teknụzụ nhicha nke bead magnet, ọ dịghịkwa mkpa iwepụ phenol chloroform na-egbu egbu n'oge usoro mwepụ. Ọ dị nchebe, ọ naghị egbu egbu ma dịkwa ngwa ngwa. Enwere ike iji ngwaahịa a ozugbo na PCR, qPCR, usoro ọgbọ nke abụọ na nnwale ndị ọzọ. Mgbe ejiri ya na ngwa mwepụta magnetic bead akpaka, enwere ike nweta mwepụta dị elu nke nucleic acid.
Ozi ngwaahịa
| Akụkụ | 48T |
| Ngwọta Lysis na Njikọta | 33 ml |
| Nkwụsị ihe ndọta magnetik | 1.1 mL |
| Ihe nchekwa ịsa ákwà A | 40 ml |
| Ihe nchekwa ịsa ákwà B | 80 ml |
| Ngwọta iwepụ ihe | 10 ml |
| Proteinase K | 1.1 mL |
Ọnọdụ Nchekwa
A ga-echekwa Proteinase K na okpomọkụ 2 ~ 8℃, a ga-echekwakwa ihe ndị ọzọ dị na ya n'ọnọdụ okpomọkụ ụlọ. Ndụ nchekwa ahụ bụ ọnwa 18.
Nkọwa Ihe Nlereanya
1. Ụdị ihe nlele ndị dị mkpa: ọbara dum, serum, plasma, mmiri cerebrospinal, swabs imi/pharyngeal, mmiri ịsa ahụ alveolar na ihe nlele ndị ọzọ.
2. Nchekwa na mbufe ihe nlele: Enwere ike iji ihe nlele mee nnwale ozugbo ma ọ bụ chekwaa ya na -70℃ ma ọ bụ karịa maka nnwale. Oge nchekwa ahụ bụ ọnwa isii. A gaghị ahapụ ifriizi na ịgbaze ugboro ugboro. A na-eji usoro oyi ebuga ihe nlele ahụ.
3. Ihe achọrọ maka ifriizi na ịgbaze: Ịkpọnwụ ngwa ngwa na ịgbaze, zere ifriizi na ịgbaze ugboro ugboro.
Ọrụ
Tupu nnwale ahụ, lelee ma mmiri mmiri na-ezo na ma enwere ike ịkwụsịtụ ihe ndọta ahụ ọzọ.
1. Bufee 200 ~ 300 μL nke ihe nlele ahụ na tube centrifuge 1.5 mL, tinye 600 μL nke Lysis na Binding Solution, 20 μL nke Proteinase K, na 20 μL nke magnet bead suspension n'usoro, wee vortex na oke ọsọ maka sekọnd 30. Kpoo ya na 50℃ maka nkeji 5 ~ 7. Ọ bụrụ na thermostat enweghị ọrụ oscillation, vortex ugboro atọ n'oge incubation, oge ọ bụla maka sekọnd 15.
2. Bufee ya na ihe ndọta 1.5 mL maka nkewa magnetik ruo mgbe ngwọta ahụ doro anya ma doo anya, tụfuokwa ngwọta ahụ.
3. Tinye 700 μL nke ihe nchekwa ihe nhicha A, vortex maka sekọnd iri iji kewaa ihe mkpuchi ihe ndọta, wee bufee ya na ihe nkwụnye magnet maka nkewa magnet ruo mgbe ngwọta ahụ doro anya, wee tụfuo ngwọta ahụ.
4. Tinye 700 μL nke ihe nchekwa ihe nhicha ahụ B, vortex ruo sekọnd iri iji kewaa ihe mkpuchi ihe ndọta ahụ, bufee ya na ihe nkwụnye magnet maka nkewa magnet ruo mgbe ngwọta ahụ doro anya, wee tụfuo ngwọta ahụ.
5. Tinye 700 μL nke ihe nchekwa ihe nhicha ahụ B ọzọ, vortex ruo sekọnd iri iji kewaa ihe mkpuchi ihe ndọta ahụ, bufee ya na ihe nkwụnye magnet maka nkewa magnet ruo mgbe ngwọta ahụ doro anya, wee tụfuo ngwọta ahụ.
6. Jiri centrifuge mee obere oge iji chịkọta ụmụ irighiri mmiri dị na mgbidi ọkpọkọ ahụ, bufee ya na ebe ndọta maka nkọwa, wee mịnye mmiri fọdụrụnụ. Kpọọ ya n'ikuku maka nkeji 3-5.
Rịba ama: Ihe fọdụrụ n'ime ethanol ga-egbochi mmeghachi omume enzyme ndị na-esote, yabụ hụ na ethanol ahụ na-ekupụ kpamkpam mgbe ọ kpọrọ nkụ. Akpọla ya ruo ogologo oge iji zere imetụta mmetụta mwepụ ahụ na-esote.
7. Tinye 40 ~ 100 μL nke mmiri Elution, vortex na oke ọsọ maka nkeji 2 ~ 3 iji gbajie ihe ndọta magnet. Kpoo ya na 60℃ maka nkeji 5, wee kụọ ya na oke ọsọ maka sekọnd 60.
8. Centrifuge ozugbo iji chịkọta mmiri mmiri dị na mkpuchi tube ahụ wee bufee ya na rack magnet ruo mgbe mkpụrụ osisi magnet ahụ ga-amịkọrọ kpamkpam. Mgbe ahụ, jiri nlezianya bufee mmiri mmiri ahụ na tube centrifuge ọhụrụ iji nweta ngwọta nucleic acid.
9. Enwere ike ịchekwa ihe ngwọta nucleic acid na -20°C maka nchekwa nwa oge na -80°C maka nchekwa ogologo oge.
Ihe ndetu
1. Ihe nchekwa dị iche iche dị na ngwa a nwere nnu guanidine. Maka nchekwa na ahụike gị, biko yi uwe nyocha na uwe aka a na-atụfu atụfu. Jiri aka gị mee ihe dịka iwu nchekwa si dị. Ekwela ka ihe nchekwa ahụ metụ akpụkpọ ahụ, anya na akpụkpọ ahụ mucous. Ọ bụrụ na ọ mee, biko jiri mmiri buru ibu saa ya ozugbo ma chọọ enyemaka ahụike.
2. Ọ bụrụ na ihe ngwọta ahụ apụta, a ga-asa ya na mmiri 30℃ ruo mgbe ihe mkpuchi ahụ ga-agbaze kpamkpam tupu ejiri ya.
3. Ọ bụrụ na ihe ndọta magnetik ahụ akpọnwụọla, ejila ya.
4. Ihe nchekwa dị iche iche dị na ngwa a nwere nnu guanidine. Ejila ọgwụ ndị na-eme ka ihe dị ọcha dị ka sodium hypochlorite gwọọ ha, ma ọ bụghị ya, gas ndị na-egbu egbu ga-apụta, a ga-ewerekwa ha dị ka ihe mkpofu ahụike.
5. Enwere ike inwe ihe ndọta magnetik fọdụrụnụ n'oge a na-agbapụ ihe. Gbalịa izere iku ume magnetik mgbe ị na-achọ ihe nlele.