Ikhithi ye-Virus DNA/RNA Extraction
Ikhithi(HC1009B) ingakhipha ngokushesha i-high-purity viral nucleic acids (DNA/RNA) kumasampula oketshezi ahlukahlukene njengegazi, i-serum, i-plasma, noketshezi lokuwasha lwe-swab, okuvumela ukucutshungulwa okuphezulu kwamasampula afanayo.Ikhithi isebenzisa ubuhlalu obuzibuthe obususelwe ku-superparamagnetic silicon obuhlukile.Ohlelweni lwebhafa oluyingqayizivele, ama-nucleic acid esikhundleni samaprotheni nokunye ukungcola akhangiswa ngamabhondi e-hydrogen nokubopha i-electrostatic.Ubuhlalu obukazibuthe obunama-nucleic acid adsorbed bugezwa ukususa amaprotheni nosawoti asele.Uma usebenzisa isivimbeli esinosawoti ophansi, ama-nucleic acid akhululwa ebuhlaluni obukazibuthe, ukuze kufezwe injongo yokuhlukana ngokushesha nokuhlanzwa kwama-nucleic acid.Yonke inqubo yokusebenza ilula, iyashesha, iphephile futhi iyasebenza, futhi ama-nucleic acid atholakele angasetshenziswa ngokuqondile ekuhloleni okuphansi komfula okufana nokubhala okuphambene, i-PCR, i-qPCR, i-RT-PCR, i-RT-qPCR, ukulandelana kwesizukulwane esilandelayo, ukuhlaziywa kwe-biochip, njll.
Izimo zokugcina
Gcina ku-15 ~ 25 ℃, futhi uhambise ekamelweni lokushisa.
Izinhlelo zokusebenza
Igazi, i-serum, i-plasma, i-swab eluent, i-tissue homogenate nokunye.
Inqubo Yokuhlola
1. Isampula ukucubungula
1.1 Kumagciwane kumasampuli awuketshezi njengegazi, i-serum, neplasma: 300μL we-supernatant esetshenziselwa ukukhipha.
2.2 Ukuze uthole amasampula e-swab: Faka amasampula e-swab kumashubhu esampula aqukethe isisombululo sokulondoloza, i-vortex iminithi elingu-1, bese uthatha i-supernatant engu-300μL ukuze uyikhiphe.
1.3 Ngamagciwane aku-tissue homogenates, izixazululo ze-tishusoak, namasampuli emvelo: Yima amasampula imizuzu engu-5 -10, bese uthatha u-300μL we-supernatant ukuze ukhishwe.
2. Ukulungiselela prepi-reagent ehlanganisiwe
Khipha ama-reagents ahlanganiswe ngaphambilini kukhithi, guqula futhi uhlanganise izikhathi ezimbalwa ukuze umise futhi ubuhlalu obuzibuthe.Nyakazisa ipuleti ngobumnene ukuze wenze ama-reagents nobuhlalu obukazibuthe bucwiliswe phansi komthombo.Sicela uqinisekise isiqondiso sepuleti futhi udabule ngokucophelela ucwecwe lwe-aluminium ovala uphawu.
Δ Gwema ukudlidliza lapho udabula ifilimu evalayo ukuze uvikele uketshezi ukuthi lungachitheki.
3. Ukusebenza kwe i-autominsimbi ye-atic
3.1 Engeza u-300μL wesampula emithonjeni Emakholomu 1 noma 7 epuleti lomthombo ojulile elingu-96 (naka indawo yokusebenza kahle yokusebenza kahle).Ivolumu yokufaka yesampula ihambisana no-100-400 μL.
3.2 Faka ipuleti lequla elijule kahle elingu-96 kusikhiphi se-nucleic acid.Gqoka imikhono yamagnetic bar, futhi uqinisekise ukuthi imboza ngokuphelele izinti kazibuthe.
3.3 Setha uhlelo ngendlela elandelayo ukuze lukhishwe ngokuzenzakalelayo:
3.4 Ngemva kokukhipha, dlulisela i-event Ikholomu 6 noma 12 ye-deep well plate engu-96 (naka indawo esebenza kahle yokusebenza kahle) uyiyise eshubhuni elihlanzekile le-Nuclease-free centrifuge.Uma ungayisebenzisi ngokushesha, sicela ugcine imikhiqizo ku -20 ℃.
Amanothi
Okokusetshenziswa kocwaningo kuphela.Ayisetshenziselwa izinqubo zokuxilonga.
1. Umkhiqizo okhishiwe yi-DNA/RNA.Ukunakwa okukhethekile kufanele kukhokhwe ukuvimbela ukuwohloka kwe-RNA nge-RNase ngesikhathi sokusebenza.Izitsha namasampula asetshenzisiwe kufanele anikezelwe.Wonke amashubhu namathiphu we-pipette kufanele abulawe amagciwane futhi angabi ne-DNase/RNase.Abasebenzi kufanele bagqoke amagilavu namamaski angenayo impushana.
2. Sicela ufunde imanuwali yeziqondiso ngokucophelela ngaphambi kokuyisebenzisa, futhi usebenze ngokuhambisana ngokuqinile nencwadi yeziqondiso.Ukucubungula isampula kufanele kwenziwe ebhentshini elihlanzekile kakhulu noma ikhabhinethi yokuphepha yebhayoloji.
3. Isistimu yokukhipha i-nucleic acid ezenzakalelayo kufanele ibulawe amagciwane nge-UV imizuzu engama-30 ngaphambi nangemva kokusetshenziswa.
4. Kungase kube khona imikhondo yobuhlalu kazibuthe obusele ku-event ngemva kokukhishwa, ngakho gwema ukulangazelela ubuhlalu obuzibuthe.Uma ubuhlalu kazibuthe bushiswa, bungasuswa ngokuma kazibuthe.
5. Uma ingekho imiyalelo ekhethekile yamaqoqo ahlukene ama-reagents, sicela ungawaxubi, futhi uqinisekise ukuthi amakhithi asetshenziswa phakathi nesikhathi sokufaneleka.
6. Lahla kahle wonke amasampula kanye ne-reagent, sula phansi futhi ubulale amagciwane kuzo zonke izindawo zokusebenza nge-ethanol engu-75%.
