Ikhithi yokukhipha i-Viral DNA/RNA
Le kit ifanele ukukhishwa ngokushesha kwe-high-purity virus DNA/RNA kusuka kumasampuli afana namaswabs e-nasopharyngeal, ama-swabs emvelo, ama-cell culture supernatants, nama-tissue homogenate supernatants.Ikhithi isuselwe kubuchwepheshe bokuhlanza ulwelwesi lwe-silica obuqeda isidingo sokusebenzisa izinyibilikisi eziphilayo ze-phenol/chloroform noma imvula yotshwala edla isikhathi ukuze kukhishwe i-viral DNA/RNA yekhwalithi ephezulu.Ama-nucleic acid atholiwe awanakho ukungcola futhi alungele ukusetshenziswa ekuhloleni okuya phansi komfula njengokulotshwa okuhlanekezelwe, i-PCR, i-RT-PCR, i-PCR yesikhathi sangempela, ukulandelana kwesizukulwane esilandelayo (NGS), kanye ne-Northern blot.
Izimo zokugcina
Gcina ku-15 ~ 25 ℃, futhi uhambise ekamelweni lokushisa
Izingxenye
| Izingxenye | 100RXNS |
| Isivimbeli VL | 50 ml |
| Isilondolozi se-RW | 120 ml |
| I-ddH2 O yamahhala ye-RNase | 6 ml |
| I-FastPure RNA Amakholomu | 100 |
| Amashubhu Okuqoqwayo (2ml) | 100 |
| Amashubhu Eqoqo Angenayo i-RNase(1 .5ml) | 100 |
I-Buffer VL:Nikeza indawo ye-lysis nokubopha.
Isivimbeli RW:Susa amaprotheni ayinsalela nokunye ukungcola.
I-ddH2O yamahhala ye-RNase:I-Elute DNA/RNA kusukela kulwelwesi kukholomu yokuphotha.
Amakholomu e-FastPure RNA:I-adsorb DNA/RNA ngokukhethekile.
Amashubhu eqoqo 2 ml:Qoqa isihlungi.
Amashubhu Eqoqo Angenayo i-RNase 1.5 ml:Qoqa i-DNA/RNA.
Izinhlelo zokusebenza
Ama-swabs e-nasopharyngeal, ama-swabs emvelo, ama-cell culture supernatants, nama-tissue homogenate supernatants.
Uzilungiselela Materials
Amathiphu epayipi angenayo i-RNase, amashubhu e-centrifuge angenayo i-RNase engu-1.5 ml, i-centrifuge, i-vortex mixer, namapayipi.
Inqubo Yokuhlola
Yenza zonke lezi zinyathelo ezilandelayo kukhabhinethi ye-biosafety.
1. Engeza u-200 μl wesampula kushubhu ye-centrifuge engenayo i-RNase (yenza ne-PBS noma i-0.9% NaCl uma kunesampula enganele), engeza u-500 μl we-Buffer VL, xuba kahle ngokuvortex amasekhondi angu-15 – 30, kanye ne-centrifuge. kafushane ukuqoqa ingxube phansi tube.
2. Faka Amakholomu e-FastPure RNA Kumashubhu Eqoqo 2 ml.Dlulisa ingxube isuka ku-Isinyathelo 1 uye kumakholomu e-FastPure RNA, i-centrifuge ku-12,000 rpm (13,400 × g) iminithi elingu-1, bese ulahla i-filtrate.
3. Engeza u-600 μl we-Buffer RW kumakholomu e-FastPure RNA, i-centrifuge ku-12,000 rpm (13,400 × g) amasekhondi angu-30, futhi ulahle isihlungi.
4. Phinda Isinyathelo sesi-3.
5. Faka i-Centrifuge ikholomu engenalutho ku-12,000 rpm (13,400 × g) imizuzu emi-2.
6. Dlulisa ngokucophelela Amakholomu e-FastPure RNA uwafake ku-RNase-free Collection Tubes entsha engu-1.5 ml (ehlinzekwe kukhithi), bese wengeza u-30 – 50 μl we-ddH2O yamahhala ye-RNase phakathi nendawo yolwelwesi ngaphandle kokuthinta ikholomu.Vumela ukuma ekamelweni lokushisa iminithi elingu-1 kanye ne-centrifuge ku-12,000 rpm (13,400 × g) iminithi elingu-1.
7. Lahla Amakholomu e-FastPure RNA.I-DNA/RNA ingasetshenziswa ngokuqondile ekuhloleni okulandelayo, noma igcinwe ku -30~ -15°C isikhathi esifushane noma -85 ~-65°C isikhathi eside.
Amanothi
Okokusetshenziswa kocwaningo kuphela.Ayisetshenziselwa izinqubo zokuxilonga.
1. Linganisa amasampula ekamelweni lokushisa kusengaphambili.
2. Amagciwane athathelwana kakhulu.Sicela uqinisekise ukuthi zonke izinyathelo zokuphepha ezidingekayo ziyathathwa ngaphambi kokuhlolwa.
3. Gwema ukuqhwaza okuphindaphindiwe nokuncibilika kwesampula, njengoba lokhu kungase kuholele ekulimazeni noma ekwehliseni isivuno se-viral DNA/RNA ekhishiwe.
4. Izinto ezizilungiselelayo zihlanganisa amathiphu epayipi angenayo i-RNase, amashubhu e-centrifuge angenayo i-RNase engu-1.5 ml, i-centrifuge, i-vortex mixer, namapayipi.
5. Lapho usebenzisa ikhithi, gqoka ijazi lelebhu, amagilavu e-latex alahlwayo, kanye nemaski alahlwayo futhi usebenzise izinto ezisebenzisekayo ezingenawo i-RNase ukuze unciphise ubungozi bokungcola kwe-RNase.
6. Yenza zonke izinyathelo ekamelweni lokushisa ngaphandle uma kuchazwe ngenye indlela.
Indlela & Ukuhamba komsebenzi
