IViral RNA Isolation Kit yokuCocwa kweViral RNA kwiPlasma, iSerum, kunye nezinye iiSampuli
Inkcazelo yeKit:
Ikati.No.RE-02011/02014
Ukucocwa kwe-RNA yentsholongwane kwiplasma, i-serum, ulwelo lomzimba olungenaziseli, amandla amakhulu enkcubeko yeeseli.
Ukwahlula ngokukhawuleza kwaye ucoce i-RNA yentsholongwane kwiisampulu ezinje ngeplasma, i-serum, ulwelo lomzimba olungenaziseli kunye nezamandla zenkcubeko yeeseli.
-Akukho mfuneko yokukhathazeka malunga nokuthotywa kwe-RNA.Ikhithi iyonke i-RNase-Free
-Elula-yonke imisebenzi igqitywe kwiqondo lokushisa
-Ukukhawuleza-ukusebenza kunokugqitywa kwimizuzu engama-20
-Isivuno esiphezulu se-RNA: IKholamu ye-RNA kuphela kunye nefomula ekhethekileyo inokucoca ngokufanelekileyo i-RNA
-Ikhuselekile-akukho reagent ephilayo esetyenzisiweyo
-Umthamo omkhulu wokusetyenzwa kwesampulu-ukuya kuthi ga kwi-200μl iisampulu zinokucutshungulwa ngexesha ngalinye.
-Umgangatho ophezulu-i-RNA ehlambulukileyo icocekile kakhulu, ayinaprotheyini kunye nobunye ukungcola, kwaye inokuhlangabezana nezicelo ezahlukeneyo zokulinga.
Iinkcazelo
Ikhithi isebenzisa ikholamu ye-spin kunye nefomula ephuhliswe yi-Foregene, enokuthi ikhuphe ngokufanelekileyo ubunyulu obuphezulu kunye nekhwalithi ephezulu ye-RNA yentsholongwane kwiisampulu ezinje ngeplasma, i-serum, i-cell-free body fluid, kunye ne-cell culture supernatant.Ikiti yongeza ngokukodwa i-Linear Acrylamide, enokuthi ibambe ngokulula ixabiso elincinci le-RNA kwiisampuli.IKholam ye-RNA Kuphela inokubopha i-RNA ngokufanelekileyo.Ikhithi inokuqhuba inani elikhulu leesampuli ngexesha elinye.
Ikhithi yonke ayinayo i-RNase, ngoko ke i-RNA ecocekileyo ayiyi kuthotywa.I-Buffer viRW1 kunye ne-Buffer viRW2 inokuqinisekisa ukuba i-viral nucleic acid efunyenweyo ayinaprotheyini, i-nuclease okanye enye into engcolileyo, enokusetyenziswa ngokuthe ngqo kwimifuniselo yebhayoloji yemolekyuli esezantsi.
Iinkcukacha
50 Preps, 200 Preps
Amacandelo ekhithi
| Umgca Acrylamide |
| Buffer viRL |
| Isithinteli viRW1, Isithinteli viRW2 |
| RNase-Free ddH2O |
| Ikholamu ye-RNA Kuphela |
| Imiyalelo |
Iimpawu&izinto eziluncedo
-Akukho mfuneko yokukhathazeka malunga nokuthotywa kwe-RNA.Ikhithi iyonke i-RNase-Free
-Elula-yonke imisebenzi igqitywe kwiqondo lokushisa
-Ukukhawuleza-ukusebenza kunokugqitywa kwimizuzu engama-20
-Isivuno esiphezulu se-RNA: IKholamu ye-RNA kuphela kunye nefomula ekhethekileyo inokucoca ngokufanelekileyo i-RNA
-Ikhuselekile-akukho reagent ephilayo esetyenzisiweyo
-Umthamo omkhulu wokusetyenzwa kwesampulu-ukuya kuthi ga kwi-200μl iisampulu zinokucutshungulwa ngexesha ngalinye.
-Umgangatho ophezulu-i-RNA ehlambulukileyo icocekile kakhulu, ayinaprotheyini kunye nobunye ukungcola, kwaye inokuhlangabezana nezicelo ezahlukeneyo zokulinga.
Iiparamitha zeKit
Isicelo seKit:
Ifanelekile ukutsalwa kunye nokucocwa kwe-RNA yentsholongwane kwiisampuli ezifana neplasma, i-serum, i-cell-free body fluid kunye ne-cell culture supernatant.
Ukuhamba komsebenzi
Iimeko zokugcina
- Ikhithi ingagcinwa iinyanga ezingama-24 kwiqondo lobushushu begumbi (15-25℃), okanye 2-8℃ ixesha elide.
-Isisombululo se-Acrylamide esine-Linear sinokugcinwa kwiqondo lokushisa leentsuku ze-7.Emva kokufumana ikhithi, nceda uthathe isisombululo se-Linear Acrylamide kwaye usigcine ku -20 ℃.
-Emva kokongeza i-Linear Acrylamide kwi-Buffer viRL, inokugcinwa kwi-2-8℃ ukuya kwi-48h.Nceda usebenzise isisombululo esitsha esilungisiweyo.
Izikhokelo zohlalutyo lweengxaki
The following is an analysis of the problems that might be encountered in the extraction of viral RNA. We wish it would be helpful to your experiment. In addition, for other experimental or technical problems other than operating instructions and problem analysis, we have dedicated technical support to help you. Contact us if you need at : 028-83361257or E-mail：Tech@foregene.com。
Akukho RNA inokukhutshwa okanye isivuno se-nucleic acid siphantsi
Ngokuqhelekileyo zininzi izinto ezichaphazela ukusebenza kakuhle kokubuyisela, ezinje: umxholo we-RNA, indlela yokusebenza, umthamo we-elution, njl.
Uhlalutyo lwezizathu eziqhelekileyo:
1.I-ice bath okanye iqondo lokushisa eliphantsi (4 ° C) i-centrifugation ngexesha lokusebenza.
Ingcebiso: Ubushushu begumbi (15-25 ° C) ukusebenza, ungaze uhlambe umkhenkce kunye nobushushu obuphantsi be-centrifuge.
2. Ukugcinwa kwesampuli engafanelekanga okanye ukugcinwa kwesampuli ixesha elide.
Ingcebiso: Gcina iisampulu kwi-80 ° C okanye umkhenkce kwinitrogen engamanzi, kwaye ugweme ukusetyenziswa okuphindaphindiweyo komkhenkce;zama ukusebenzisa iisampulu ezisanda kuqokelelwa ukutsalwa kwe-RNA.
3.Isampula enganelanga i-lysis
Isincomo: Nceda uqinisekise ukuba isampuli kunye nesisombululo esisebenzayo (i-Linear Acrylamide) ixutywe ngokucokisekileyo kwaye ifakwe kwi-10 min kwindawo yokushisa (15-25 ° C)
4.I-eluent yongezwa ngokungalunganga
Ingcebiso: Qinisekisa ukuba i-ddH2O ye-RNase-Free yongezwa kumbindi wenwebu yekholamu yokucoca.
5.Umthamo ongafanelekanga we-ethanol ye-anhydrous kwi-Buffer viRW2
Ingcebiso: Nceda ulandele imiyalelo, yongeza umthamo ochanekileyo we-ethanol ye-anhydrous kwi-Buffer viRW2 kwaye uyixube kakuhle ngaphambi kokusebenzisa ikhithi.
6.Ukusetyenziswa kwesampulu engafanelekanga.
Iingcebiso: 200µl yesampulu nge-500μl ye-Buffer viRL.Umthamo wesampulu ogqithisileyo uya kukhokelela ekunciphiseni izinga lokutsalwa kwe-RNA.
7.Umthamo we-elution ongafanelekanga okanye ukukhutshwa okungaphelelanga.
Isiphakamiso: Umthamo ocacileyo wekholamu yokucoca yi-30-50μl;ukuba isiphumo asanelisi, kuyacetyiswa ukuba udibanise i-RNase-Free ddH eshushu ngaphambili.2O kunye nokwandisa ixesha lokubeka kwindawo yokushisa, njenge-5-10min
8.Ikholamu yokucoca inentsalela ye-ethanol emva kokuhlanjululwa kwi-Buffer viRW2.
Isiphakamiso: Ukuba i-ethanol isala emva kokuhlanjululwa kwi-Buffer viRW2 kunye ne-centrifugation ye-tube engenanto ye-2min, ikholamu yokucoca inokushiywa kwindawo yokushisa ye-5min emva kwe-tube centrifugation engenanto ukususa ngokupheleleyo i-ethanol eseleyo.
Ukonakaliswa kweemolekyuli ze-RNA ezihlambulukileyo
Umgangatho we-RNA ecocekileyo inxulumene nezinto ezifana nokugcinwa kwesampulu, ukungcoliseka kwe-RNase, kunye nokusebenza.
Uhlalutyo lwezizathu eziqhelekileyo:
1.Iisampulu eziqokelelweyo azigcinwanga ngexesha.
Ingcebiso: Ukuba isampulu ayisetyenziswanga ngexesha emva kokuqokelela, nceda uyigcine ku -80 ℃ okanye initrogen engamanzi ngoko nangoko.Xa kutsalwa iimolekyuli ze-RNA, zama ukusebenzisa iisampulu ezisanda kuqokelelwa xa kunokwenzeka.
2.Iisampulu eziqokelelweyo zazikhenkceza kwaye zinyibilike ngokuphindaphindiweyo.
Isiphakamiso: Gwema ukukhenkceza ngokuphindaphindiweyo kunye nokunyibilika (kungekho ngaphezu kweyodwa) ngexesha lokuqokelela isampuli kunye nokugcinwa, ngaphandle koko isivuno se-nucleic acid siya kuncipha.
3.I-RNase yaziswa kwigumbi lokusebenza okanye akukho ziglavu ezilahlwayo, iimaski, njl.njl.
Ingcebiso: Ukutsalwa kovavanyo lweemolekyuli ze-RNA kwenziwa ngcono kwigumbi lokusebenza elahlukileyo le-RNA, kwaye itafile yovavanyo iyacocwa phambi kovavanyo.Nxiba iiglavu ezilahlwayo kunye nemaski ngexesha lovavanyo ukunqanda ukuthotywa kwe-RNA okubangelwa kukwaziswa kwe-RNase.
I-4.I-reagent ingcolisekile yi-RNase ngexesha lokusetyenziswa.
Ingcebiso: Faka enye iViral RNA Isolation Kit kwimifuniselo ehambelanayo.
5.Ungcoliseko lwe-RNase yeetyhubhu ze-centrifuge, iingcebiso ze-pipette, njl. Ingcebiso: Qinisekisa ukuba iityhubhu ze-centrifuge, iingcebiso ze-pipette, kunye neepayipi zonke azina-RNase.
Iimolekyuli ze-RNA ezisulungekileyo zichaphazele iimvavanyo ezisezantsi
Iimolekyuli ze-RNA ezihlanjululwe yikholomu yokucoca ziya kuchaphazela iimvavanyo ezisezantsi ukuba kukho ii-ion zetyuwa ezininzi okanye iiprotheni, ezinje: ukuguqulelwa umva, i-Northern Blot, njl.
1.Kukho iiyoni zetyuwa ezishiyekileyo kwiimolekyuli ze-RNA eluted.
Isincomo: Qinisekisa ukuba umthamo ochanekileyo we-ethanol ye-anhydrous yongezwe kwi-Buffer viRW2, kwaye uhlambe ikholamu yokucoca kabini ngokwesantya esichanekileyo se-centrifugation kwimiyalelo yokusebenza; Ukuba kusekho i-ion yetyuwa eseleyo, unokongeza i-Buffer viRW2 kwikholamu yokucoca, kwaye uyishiye kwindawo yokushisa ye-5min.Emva koko yenza i-centrifugation ukususa i-ion yetyuwa ngowona mgangatho mkhulu
2.Kukho i-ethanol eshiyekileyo kwiimolekyuli ze-RNA
Isiphakamiso: xa uqinisekisa ukuba iikholomu zokucoca zihlanjululwe yi-Buffer viRW2, yenza i-centrifugation ye-tube engenanto ngokwesantya se-centrifugal kwimiyalelo yokusebenza.Ukuba kusekho i-ethanol eseleyo, inokushiywa imizuzu emi-5 kwiqondo lobushushu begumbi emva kokuba i-centrifugation yetyhubhu engenanto ukususa i-ethanol eseleyo ngowona mlinganiselo mkhulu.
Izikhokelo zoMyalelo:
I-Viral RNA Isolation Kit Manual Manual
