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iphepha_ibhena

ISeli iyonke yeRNA Isolation Kit Iyonke iRNA Isolation IiKits zokucoca ukusuka kwiseli

Inkcazelo yeKit:

Isulungekiswe kakhulu kwaye ikumgangatho ophezulu we-RNA iyonke inokufumaneka kwiiseli ezahlukeneyo zenkcubeko kwimizuzu eli-11.

RNase-Free

Isebenza kubushushu begumbi (15-25℃)

NgeKholamu yokucoca yeDNA

Isivuno esiphezulu se-RNA

Ukukhawuleza:Gqiba ukutsalwa ngemizuzu eyi-11

Ukhuseleko:Ayikho imichiza ye-Organic

amandla angaphambili


  • :
  • Iinkcukacha zeMveliso

    Iithegi zeMveliso

    FAQ

    Inkcazo

    Le kit isebenzisa ikholomu ye-spin kunye nefomula ephuhliswe yi-Foregene, enokuthi ikhuphe ngokufanelekileyo ubunyulu obuphezulu kunye nomgangatho ophezulu we-RNA kwiiseli ezikhuliswe kwi-96, i-24, i-12, kunye ne-6-well-plates.

    Ikiti ibonelela ngeKholamu ye-DNA-Cleaning esebenzayo, ekwazi ukwahlula ngokulula i-supernatant kunye ne-cell lysate, ibophe kwaye isuse i-genomic DNA.Umsebenzi ulula kwaye wonga ixesha.

    Tyena iKholamu ye-RNA kuphela inokubopha ngokufanelekileyo i-RNA ngefomula eyodwa.Inani elikhulu leisampulu linokucutshungulwa ngaxeshanye.

    Amacandelo ekhithi

    Ukwakhiwa kweKit RE-03111 RE-03114
    50 T 200 T
    Isithinteli cRL1* 25 ml 100 ml
    Isithinteli cRL2 15 ml 60 ml
    Isithinteli RW1* 25 ml 100 ml
    Isithinteli RW2 24 ml 96 ml
    RNase-Free ddH2O 10 ml 40 ml
    Ikholamu ye-RNA Kuphela 50 200
    Ikholamu yokucoca iDNA 50 200
    Umyalelo 1 1

    *Nceda unxibe iiglavu kwaye uthathe amanyathelo okukhusela ngexesha lomsebenzi njengoko i-Buffer cRL1 kunye ne-Buffer RW1 ziqulethe iityuwa ze-chaotropic ezicaphukisayo.

    Iimpawu&izinto eziluncedo

    ■ Yonke le nkqubo iqhutywa kubushushu begumbi (15-25℃), ngaphandle kokuhlamba komkhenkce kunye nobushushu obuphantsi be-centrifugation.
    ■ Ikhithi iyonke i-RNase-Free, akukho mfuneko yakuzikhathaza ngokonakala kwe-RNA.
    ∎ IKholam yokucoca i-DNA ibophelela ngokukodwa i-DNA, ukuze ikhithi ikwazi ukususa ungcoliseko lwe-DNA ye-genomic ngaphandle kokongeza i-DNase.
    ■ Isivuno esiphezulu se-RNA: IKholamu ye-RNA kuphela kunye nefomula ekhethekileyo inokucoca ngokufanelekileyo i-RNA.
    ■ Isantya esikhawulezayo: kulula ukusebenza kwaye sinokugqitywa ngemizuzu eyi-11.
    ■ Ukhuseleko: Akukho sixhobo sokwenza izinto eziphilayo esifunekayo.
    ∎ Umgangatho ophezulu: I-RNA ecociweyo yeyobunyulu obuphezulu, ayinaprotheyini kunye nobunye ukungcola, kwaye inokumelana nemifuniselo eyahlukeneyo elandelayo.

    iingenelo ze-foregene RNA Isolation kit

    Isicelo seKit

    Ifanelekile ukutsalwa kunye nokuhlanjululwa kwe-RNA epheleleyo kwiiseli ezikhuliswe kwii-96, 24, 12, kunye ne-6-well-plates.

    Ukuhamba komsebenzi

    iseli iyonke RNA

    Umzobo

    ISeli iyonke yeRNA yokuBekwa kweKhiti yokuSebenza1

    Umzobo webhetri yejeli ye-agarose yeCell Total RNA Isolation Kit yaphatha amanani angasentla ahlukeneyo eeseli, 20μl umthamo we-elution, thatha i-2μl esulungekisiweyo iyonke ye-RNA 1%.

    Ukugcinwa kunye nobomi beshelufu

    Ikhithi ingagcinwa iinyanga ezili-12 kwiqondo lobushushu begumbi (15–25 ℃) okanye 2–8 ℃ ixesha elide (iinyanga ezingama-24).

    I-Buffer cRL1 inokugcinwa kwi-4 ℃ kwinyanga ye-1 emva kokongeza i-2-hydroxy-1-ethanethiol (ukhetho).


  • Ngaphambili:
  • Okulandelayo:

  • I-RNA ayikhutshwa okanye izivuno ze-RNA ziphantsi

    Kuhlala kukho izinto ezahlukeneyo ezichaphazela ukusebenza kakuhle kokubuyisela, njengale: umxholo we-RNA we-tissue, indlela yokusebenza, umthamo we-lution, njl.

    1. Ibhafu ye-ice okanye i-cryogenic (4 ° C) i-centrifugation yenziwa ngexesha lokusebenza.

    Isincomo: Sebenza kwiqondo lokushisa (15-25 ° C) kuyo yonke inkqubo, musa ukuhlamba i-ice kunye ne-centrifuge kumaqondo aphantsi.

    2. Ukugcinwa kwesampulu engafanelekanga okanye ixesha lokugcina isampuli.

    Isindululo: Gcina iisampulu kwi -80 °C okanye umkhenkce kwinitrogen engamanzi kwaye uthintele ukusetyenziswa okuphindaphindiweyo komkhenkce;zama ukusebenzisa izicubu ezitsha okanye iiseli ezikhuliswayo zokutsalwa kwe-RNA.

    3. I-lysis yesampuli eyaneleyo.

    Isincomo: Xa i-homogenizing tissue, qinisekisa ukuba i-tissue i-homogenized ngokwaneleyo kwaye iiseli zethishu zahlulwe ngokwaneleyo ukucacisa ukukhululwa kwe-RNA.

    4. I-eluent ayidityaniswa ngokuchanekileyo.

    Ingcebiso: Qinisekisa ukuba i-RNase-Free ddH2I-O yongezwa kwi-dropwise phakathi kwe-membrane yekholomu yokucoca.

    5. Umthamo ochanekileyo we-absolute ethanol awuzange wongezwe kwi-Buffer RL2 okanye kwi-Buffer RW2.

    Ingcebiso: Landela imiyalelo, yongeza umthamo ochanekileyo we-ethanol epheleleyo kwi-Buffer RL2 kunye ne-Buffer RW2 kwaye udibanise kakuhle phambi kokusebenzisa ikhithi.

    6. Idosi yesampula yezicubu ayifanelekanga.

    Isincomo: Sebenzisa i-10-20 mg yezicubu okanye (1-5) × 106iiseli nge-500 μl buffer RL1, njengoko ukusetyenziswa kwethishu ngokugqithisileyo kunokubangela ukutsalwa kwe-RNA okucuthiweyo.

    7. Umthamo ongafanelekanga okanye i-elution engaphelelanga.

    Isincomo: Umthamo we-elution wekholamu yokucoca yi-50-200 μl;ukuba isiphumo se-elution asanelisi, kuyacetyiswa ukuba kwandiswe ixesha lokubeka ubushushu begumbi emva kokongeza i-RNase-Free ddH efudunyeziweyo.2O, umz. kwi-5-10 min.

    8.Ikholamu yokucoca inentsalela ye-ethanol emva kokuhlamba kwe-Buffer RW2.

    Isincomo: Ukuba kukho intsalela ye-ethanol emva kokuhlamba kwe-Buffer RW2, i-tube engenanto ye-centrifugation ye-1min, ixesha le-tube engenanto ye-centrifugation operation inokunyuswa ukuya kwi-2min, okanye ikholomu yokucoca inokubekwa kwindawo yokushisa ye-5 min ukususa ngokwaneleyo intsalela. i-ethanol.

    I-RNA ecocekileyo ithotywa

    Umgangatho we-RNA ecocekileyo inxulumene nezinto ezifana nokugcinwa kwesampulu, ukungcoliswa kwe-RNase, kunye nokukhwabanisa, njl.

    1. Iisampulu zezicubu azigcinwanga ngexesha.

    Isincomo: Ukuba iisampulu zethishu okanye iiseli azisetyenziswanga ngexesha elifanelekileyo emva kokuqokelelwa, ngoko nangoko gcina i-cryopreserve kwi -80 °C okanye initrogen engamanzi.Ukukhupha i-RNA, sebenzisa ithishu esanda kuthathwa okanye isampulu yeseli xa kunokwenzeka.

    2. Ukunyibilikiswa okuphindaphindiweyo kweesampuli zethishu.

    Isincomo: Xa ugcina iisampulu zethishu, kungcono ukuba uzinqumle zibe ngamaqhekeza amancinci ukuze zigcinwe, kwaye ususe esinye seziqwenga xa uzisebenzisa ukuphepha ukunyibilika okuphindaphindiweyo kwesampulu kunye nokuthotywa kwe-RNA.

    3. I-RNase yaziswa okanye inganxibi iiglavu ezilahlwayo, iimaski, njl njl ngexesha lokusebenza.

    Isincomo: Imifuniselo yokutsalwa kwe-RNA yenziwa ngcono kumagumbi okukhohlisa e-RNA kwaye itafile iyacinywa phambi kovavanyo.

    Nxiba iiglavu ezilahlwayo kunye nemaski ngexesha lovavanyo lokunciphisa ukuthotywa kwe-RNA okubangelwa kukwaziswa kwe-RNase.

    4. Ii-reagents zingcolisekile nge-RNase ngexesha lokusetyenziswa.

    Ingcebiso: Faka endaweni entsha yeSilwanyana esipheleleyo se-RNA ye-Isolation Kit ukulungiselela imifuniselo ehambelanayo.

    5. Iityhubhu ze-centrifuge, iingcebiso, njl.njl ezisetyenziswa kwi-RNA manipulation zingcolile nge-RNase.

    Ingcebiso: Qinisekisa ukuba iityhubhu ze-centrifuge, iingcebiso, iipipette, njl.njl ezisetyenziswa kwi-RNA extraction azinayo yonke i-RNase.

    I-RNA esulungekileyo ichaphazela imifuniselo esezantsi

    I-RNA ihlanjululwe yikholomu yokucoca, ukuba iiyoni zetyuwa, umxholo weprotheyini mkhulu kakhulu uya kuchaphazela umfuniselo osezantsi, onje: uguqulelo lombhalo, Northern Blot et al.

    1. I-RNA ekhutshiweyo ineentsalela ze-ion yetyuwa.

    Isincomo: Qinisekisa ukuba umthamo ochanekileyo we-ethanol wongezwe kwi-Buffer RW2 kwaye wenze i-2 ikholamu yokuhlanjululwa kwi-speed ye-centrifugal eboniswe ekusebenzeni;ukuba kukho nayiphi na intsalela ye-ion yetyuwa, shiya ikholomu yokucoca kwi-Buffer RW2 ye-5 min kwiqondo lokushisa elingaphakathi kwaye wenze i-centrifugation ukwandisa ukukhutshwa kwetyuwa.

    2. Intsalela ye-Ethanol kwi-RNA elutioned.

    Isincomo: Qinisekisa ukuba emva kokuhlamba i-buffer RW2, yenza i-tube engenanto i-centrifugation operation kwisantya se-centrifugation esibonakaliswe ukusebenza, ukwandisa ixesha le-tube engenanto ye-centrifugation operation ukuya kwi-2 min ukuba kusekho i-ethanol intsalela, okanye uyishiye kwindawo yokushisa kwe-5. m

    Bhala umyalezo wakho apha kwaye uwuthumele kuthi