I-Blood RNA Isolation Kit
Inkcazelo yeKit:
Ikati.No.RE-04011/04013
Ukucocwa ngokupheleleyo kwe-RNA kwigazi elipheleleyo 104 ≤ Iiseli zegazi eziMhlophe ≤ 107
Ukwahlula ngokukhawuleza kwaye ucoce i-RNA yegazi kwiiseli ezimhlophe zegazi.
-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 ithatha ikholamu ye-spin kunye nefomula ephuhliswe yinkampani yethu, enokuthi ikhuphe ngokufanelekileyo ubunyulu obuphezulu kunye nekhwalithi ephezulu ye-RNA kwigazi elipheleleyo le-anticoagulated.Ikiti ibonelela nge-lysate yegazi elibomvu (i-Buffer RCL), enokuthi ngokukhawuleza kwaye isebenze ngokufanelekileyo iiseli ezibomvu zegazi kunye nokugcina iiseli ezimhlophe zegazi.I-DNA-Cleaning Colunm esebenzayo inokwahlula ngokulula i-supernatant kunye ne-cell lysates kunye ne-adsorb kwaye isuse i-DNA ye-genomic.Umsebenzi ulula kwaye wonga ixesha;IKholamu ye-RNA kuphela inokubopha ngokufanelekileyo i-RNA, kunye nefomula ekhethekileyo, inokuqhuba inani elikhulu leesampuli ngexesha elinye.
Yonke inkqubo ye-RNase-Free yenza i-RNA ekhutshiweyo ingonakaliseki;I-Buffer RW1 kunye ne-Buffer RW2 inkqubo yokuhlamba i-buffer yenza i-RNA efunyenweyo ingabi naprotheyini, i-DNA, i-ion kunye nongcoliseko lwe-organic compound.
Imixholo yeKit
| IGazi iyonke ye-RNA Isolation Kit | ||
| Ukwakhiwa kweKit | RE-04011 | RE-04013 |
| 50 amaxesha | 200 amaxesha | |
| Isithinteli RCL (10×) | 52.5 ml | 210mL |
| Isithinteli BRL1* | 30mL | 120mL |
| Isithinteli BRL2 | 18mL | 66mL |
| Isithinteli RW1* | 25mL | 100mL |
| Isithinteli RW2 | 24mL | 96mL |
| RNase-Free ddH2 O | 10mL | 40mL |
| IKholam ye-RNA kuphela | 50 iiseti | 200 iiseti |
| Ikholamu yokucoca iDNA | 50 iiseti | 200 iiseti |
| incwadi yesandla | Ikopi enye | Ikopi enye |
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 iyonke kwigazi le-mammalian elipheleleyo.
Ukuhamba komsebenzi
Iimeko zokugcina
I-Buffer RCL (10 ×) kufuneka igcinwe kwi-2-8 ℃;amanye amacandelo ekhithi angagcinwa kwiqondo lokushisa (15-25 ℃) phantsi kweemeko ezomileyo, kwaye zingagcinwa iinyanga ezili-12.I-Buffer BRL1 ingagcinwa kwi-4 ℃ kwinyanga eyi-1 emva kokongeza i-β-mercaptoethanol (ukhetho) .
Qaphela: Ukuba igcinwe kwiqondo lobushushu eliphantsi, isisombululo sithande ukuba nemvula.Qinisekisa ukuba ubeke isisombululo kwikiti kwiqondo lokushisa kwegumbi ixesha elithile ngaphambi kokusetyenziswa.Ukuba kuyimfuneko, yitshise kwindawo yokuhlambela amanzi angama-37 ° C imizuzu eyi-10 ukutshabalalisa i-precipitate, kwaye udibanise kakuhle ngaphambi kokusetyenziswa.
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
