Viral DNA / RNA Extraction Kit
Le khithi ifanelekile ukutsalwa ngokukhawuleza kwe-DNA / i-RNA yentsholongwane ephezulu kwiisampuli ezifana ne-nasopharyngeal swabs, i-swabs yendalo, i-cell culture supernatants, kunye ne-tissue homogenate supernatants.Ikiti isekelwe kwi-silica membrane yokucoca iteknoloji ephelisa isidingo sokusebenzisa i-phenol / i-chloroform i-solvents ye-organic okanye i-alcohol precipitation echitha ixesha lokukhupha i-viral DNA / RNA yomgangatho ophezulu.I-nucleic acids efunyenweyo ayinayo ukungcola kwaye ilungele ukusetyenziswa kwiimvavanyo ezisezantsi ezifana ne-reverse transcription, i-PCR, i-RT-PCR, i-PCR yexesha langempela, ukulandelelana kwesizukulwana esilandelayo (NGS), kunye ne-Northern blot.
Iimeko zokugcina
Gcina kwi-15 ~ 25 ℃, kunye nokuthutha kwiqondo lokushisa
Amacandelo
| Amacandelo | 100RXNS |
| Buffer VL | 50 ml |
| Buffer RW | 120 ml |
| RNase-free ddH2 O | 6 ml |
| IiKholamu zeRNA eziPure | 100 |
| Imibhobho yoQokelelo (2ml) | 100 |
| Iityhubhu zokuQokelelwa zasimahla ze-RNase(1 .5ml) | 100 |
Isithinteli VL:Ukubonelela ngobume be-lysis kunye nokubopha.
Isithinteli RW:Susa iiproteni ezishiyekileyo kunye nobunye ukungcola.
RNase-free ddH2O:Elute DNA/RNA ukusuka inwebu kwikholamu spin.
Iintsika zeRNA eziPure:Ngokukodwa adsorb DNA/RNA.
Imibhobho yoQokelelo 2 ml:Qokelela isihluzo.
I-RNase-free Collection Iityhubhu 1.5 ml:Qokelela i-DNA/RNA.
Usetyenziso
I-Nasopharyngeal swabs, i-swabs yokusingqongileyo, ii-supernatants zenkcubeko yeeseli, kunye ne-tissue homogenate supernatants.
Uzilungiselele Materiials
Iingcebiso ze-RNase-free pipette, i-1.5 ml ye-RNase-free centrifuge tubes, i-centrifuge, i-vortex mixer, kunye neepayipi.
Inkqubo yoVavanyo
Yenza onke la manyathelo alandelayo kwikhabhinethi ye-biosafety.
1. Yongeza i-200 μl yesampulu kwi-tube ye-centrifuge ye-RNase-free (yenza nge-PBS okanye i-0.9% ye-NaCl kwimeko yesampuli engonelanga), yongeza i-500 μl ye-Buffer VL, xuba kakuhle nge-vortexing kwi-15 - 30 sec, kunye ne-centrifuge. ngokufutshane ukuqokelela umxube ezantsi kombhobho.
2. Beka iiKholam eziFastPure zeRNA kwiiThubhu zokuQokelelwa kwe-2 ml.Dlulisa umxube ukusuka kwiNyathelo 1 ukuya kwi-FastPure RNA Columns, i-centrifuge kwi-12,000 rpm (13,400 × g) kwi-1 min, kwaye ulahle i-filtrate.
3. Yongeza i-600 μl ye-Buffer RW kwi-FastPure RNA Columns, i-centrifuge kwi-12,000 rpm (13,400 × g) ye-30 sec, kwaye ulahle i-filtrate.
4. Phinda iNyathelo lesi-3.
5. I-Centrifuge ikholomu engenanto kwi-12,000 rpm (13,400 × g) kwi-2 min.
6. Dlulisa ngononophelo iiKholamu ze-RNA ze-FastPure kwi-RNase-free Collection Tubes entsha ye-1.5 ml (ebonelelwe kwikhithi), kwaye wongeze i-30 - 50 μl ye-ddH2O ye-RNase-free kumbindi we-membrane ngaphandle kokuchukumisa ikholamu.Vumela ukuma kwindawo yokushisa kwe-1 min kunye ne-centrifuge kwi-12,000 rpm (13,400 × g) kwi-1 min.
7. Lahla iiKholamu ze-FastPure RNA.I-DNA / i-RNA ingasetyenziselwa ngokuthe ngqo kwiimvavanyo ezilandelayo, okanye igcinwe kwi -30 ~ -15 ° C ixesha elifutshane okanye -85 ~-65 ° C ixesha elide.
Amanqaku
Isetyenziselwa uphando kuphela.Ayisetyenziswa kwiinkqubo zokuxilonga.
1. Lungisa iisampuli kwiqondo lobushushu begumbi kwangaphambili.
2. Iintsholongwane ziyosulela kakhulu.Nceda uqinisekise ukuba onke amanyathelo okhuseleko ayimfuneko athathwa phambi kovavanyo.
3. Gwema ukukhenkceza ngokuphindaphindiweyo kunye nokunyibilika kwesampuli, njengoko oku kunokukhokelela ekuthotyweni okanye ukunciphisa isivuno se-viral ekhishwe i-DNA / RNA.
4. Izixhobo ezizilungiseleleyo ziquka iingcebiso ze-pipette ze-RNase, i-1.5 ml ye-RNase-free centrifuge tubes, i-centrifuge, i-vortex mixer, kunye ne-pipettes.
5. Xa usebenzisa ikhithi, nxiba idyasi yelebhu, iiglavu zelatex ezilahlwayo, kunye nemaski enokulahlwa kwaye usebenzise izinto ezisetyenziswayo ezingenazo i-RNase ukunciphisa umngcipheko wokungcoliseka kwe-RNase.
6. Yenza onke amanyathelo kwiqondo lokushisa ngaphandle kokuba kuchazwe ngenye indlela.
Inkqubo kunye nokuhamba komsebenzi
