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Ikhithi Yokuzihlukanisa Yeseli Ye-RNA

Incazelo yekhithi:

I-RNase-Free

Isebenza ekamelweni lokushisa (15-25 ℃)

Ngekholomu Yokuhlanza I-DNA

Inani eliphakeme kakhulu lama-RNA

Ukushesha:Qeda ukukhipha emizuzwini eyi-11

Ukuphepha:Ayikho i-Organic chemical

foregene strength


  • :
  • Imininingwane Yomkhiqizo

    Omaka bomkhiqizo

    FAQ

    Incazelo

    Le khithi isebenzisa ikholomu yokuphotha kanye nefomula eyakhiwe yi-Foregene, engakhipha ngempumelelo i-RNA ephezulu nekhwalithi ephezulu kumaseli akhuliswe ngamapuleti emithombo engu-96, 24, 12, kanye ne-6.

    Ikhithi inikeza Ikholomu Yokuhlanza Ye-DNA esebenza kahle, engahlukanisa kalula i-supernatant ne-cell lysate, ibophe futhi isuse i-genomic DNA.Ukusebenza kulula futhi konga isikhathi.

    TIkholomu ye-RNA kuphela ingabopha i-RNA ngokuphumelelayo ngefomula eyingqayizivele.Inani elikhulu lamasampula lingacutshungulwa kanyekanye.

    Izingxenye zekhithi

    Ukwakhiwa kwekhithi RE-03111 RE-03114
    50 T 200 T
    I-Buffer cRL1* 25 ml 100 ml
    Gcina i-cRL2 15 ml 60 ml
    Isilondolozi RW1* 25 ml 100 ml
    Isilondolozi se-RW2 24 ml 96 ml
    I-RNase-Free ddH2O 10 ml 40 ml
    Ikholomu Ye-RNA Kuphela 50 200
    Ikholomu Yokuhlanza I-DNA 50 200
    Isiyalezo 1 1

    *Sicela ugqoke amagilavu ​​futhi uthathe izinyathelo zokuzivikela ngesikhathi sokusebenza njengoba i-Buffer cRL1 ne-Buffer RW1 iqukethe usawoti we-chaotropic ocasulayo.

    Izici&izinzuzo

    ■ Yonke inqubo iqhutshwa ekamelweni lokushisa (15-25 ℃), ngaphandle kokugeza kweqhwa kanye nokushisa okuphansi kwe-centrifugation.
    ■ Yonke ikhithi ayinayo i-RNase-Free, asikho isidingo sokukhathazeka ngokuwohloka kwe-RNA.
    ■ Ikholomu Yokuhlanza I-DNA ibopha ngokukhethekile i-DNA, ukuze ikhithi isuse ukungcoliswa kwe-DNA ye-genomic ngaphandle kokwengeza i-DNase eyengeziwe.
    ■ Isivuno esikhulu se-RNA: Ikholomu ye-RNA kuphela kanye nefomula eyingqayizivele ingahlanza i-RNA ngempumelelo.
    ■ Isivinini esisheshayo: kulula ukusebenza futhi singaqedwa ngemizuzu eyi-11.
    ■ Ukuphepha: Ayikho i-organic reagent edingekayo.
    ■ Ikhwalithi ephezulu: I-RNA ehlanziwe ihlanzekile kakhulu, ayinamaprotheni nokunye ukungcola, futhi ingahlangabezana nokuhlolwa okuhlukahlukene okulandelayo.

    advantages of foregene RNA Isolation kit

    Isicelo sekhithi

    Ilungele ukukhishwa nokuhlanzwa kwe-RNA ephelele kumaseli akhulisiwe kumapuleti angama-96, 24, 12, kanye namapuleti angu-6.

    Ukugeleza komsebenzi

    cell total RNA

    Umdwebo

    Cell Total RNA Isolation Kit Work Flow1

    Umdwebo webhethri yejeli ye-agarose ye-Cell Total RNA Isolation Kit uphathe izinombolo ezingenhla ezihlukene zamaseli, ukukhishwa kwevolumu engu-20μl, thatha u-2μl isamba esihlanziwe se-RNA 1%.

    Isitoreji nempilo yeshelufu

    Ikhithi ingagcinwa izinyanga eziyi-12 ekamelweni lokushisa (15-25 ℃) noma 2-8 ℃ isikhathi eside (izinyanga ezingama-24).

    I-Buffer cRL1 ingagcinwa ku-4 ℃ inyanga engu-1 ngemva kokwengeza i-2-hydroxy-1-ethanethiol(ongakukhetha).


  • Okwedlule:
  • Olandelayo:

  • I-RNA ayikhishiwe noma isivuno se-RNA siphansi

    Kuvame ukuba nezinto ezihlukahlukene ezithinta ukusebenza kahle kokululama, njengalezi: okuqukethwe kwesampula yezicubu ze-RNA, indlela yokusebenza, ivolumu ye-elution, njll.

    1. Ukugeza kweqhwa noma i-cryogenic (4 °C) i-centrifugation yenziwa ngesikhathi sokusebenza.

    Okutuswayo: Sebenzisa ekamelweni lokushisa (15-25 ° C) kuyo yonke inqubo, ungabhavi eqhweni kanye ne-centrifuge emazingeni okushisa aphansi.

    2. Ukulondolozwa kwesampula okungalungile noma isikhathi sokugcina isampula eseqile.

    Okutuswayo: Gcina amasampula ku--80 °C noma ufrize ku-nitrogen ewuketshezi futhi ugweme ukusetshenziswa okuphindaphindiwe kokuqhwaza;zama ukusebenzisa izicubu ezintsha noma amaseli akhulisiwe ukuze kukhishwe i-RNA.

    3. Isampula yesampula enganele.

    Okutuswayo: Uma wenza izicubu zibe homogenizing, qinisekisa ukuthi izicubu zihlanganiswe ngokwanele nokuthi amaseli ezicubu ahlukaniswe ngokwanele ukuchaza ukukhululwa kwe-RNA.

    4. I-eluent ayengezwanga ngendlela efanele.

    Okunconyiwe: Qinisekisa ukuthi i-RNase-Free ddH2U-O wengezwe ngokudonsela phansi phakathi kwe-membrane yekholomu yokuhlanza.

    5. Ivolumu elungile ye-ethanol ephelele ayizange yengezwe ku-Buffer RL2 noma ku-Buffer RW2.

    Isincomo: Landela imiyalelo, engeza ivolumu efanele ye-ethanol ephelele ku-Buffer RL2 ne-Buffer RW2 bese uxuba kahle ngaphambi kokusebenzisa ikhithi.

    6. Umthamo wesampula yezicubu awufanelekile.

    Isincomo: Sebenzisa i-10-20 mg yezicubu noma (1-5) × 106amaseli nge-500 μl buffer RL1, njengoba ukusetshenziswa kwezicubu ngokweqile kungaholela ekukhishweni kwe-RNA okuncishisiwe.

    7. Ivolumu ye-elution engafanele noma ukungezwani okuphelele.

    Okutuswayo: Umthamo we-elution wekholomu yokuhlanza ungama-50-200 μl;uma umphumela we-elution ungagculisi, kuyanconywa ukuthi kunwetshwe isikhathi sokubeka izinga lokushisa legumbi ngemva kokwengeza i-ddH eshisiwe ye-RNase-Free2O, isb imizuzu emi-5-10.

    8.Ikholomu yokuhlanza inensalela ye-ethanol ngemva kokugeza kwe-Buffer RW2.

    Isincomo: Uma kunezinsalela ze-ethanol ngemva kokugeza kwe-Buffer RW2, i-tube centrifugation engenalutho ye-1min, isikhathi sokusebenza kwe-tube centrifugation esingenalutho singanyuswa sibe yi-2min, noma ikholomu yokuhlanza ingabekwa ekamelweni lokushisa imizuzu engu-5 ukuze kususwe ngokwanele okusele. i-ethanol.

    I-RNA ehlanzekile yehlisiwe

    Ikhwalithi ye-RNA ehlanzekile ihlobene nezici ezifana nokulondolozwa kwesampula, ukungcoliswa kwe-RNase, nokukhohlisa, njll.

    1. Amasampula ezicubu awagcinwa ngesikhathi.

    Okutuswayo: Uma amasampula ezicubu noma amaseli engasetshenziswa ngesikhathi esifanele ngemva kokuqoqwa, londoloza ngokushesha i-cryopreserve ku- -80 °C noma i-nitrogen ewuketshezi.Ukuze ukhiphe i-RNA, sebenzisa ithishu esanda kuthathwa noma isampula yeseli noma nini lapho kunokwenzeka.

    2. Ukuqhwaza okuphindaphindiwe kwamasampula ezicubu.

    Okutuswayo: Uma ugcina amasampula ezicubu, kuhle kakhulu ukuwasika abe izingcezu ezincane ukuze alondolozwe, bese ususa isiqeshana esisodwa lapho uzisebenzisa ukugwema ukuncibilika okubandayo kwesampula kanye nokonakala kwe-RNA.

    3. I-RNase yethulwa noma ayigqokile amagilavu ​​alahlwayo, imaski, njll. phakathi nokuhlinzwa.

    Okunconyiwe: Ukuhlolwa kokukhipha i-RNA kwenziwa kangcono kakhulu emagumbini okukhohlisa e-RNA ahlukene futhi ithebula liyasulwa ngaphambi kokuhlolwa.

    Gqoka amagilavu ​​alahlwayo kanye nemaski ngesikhathi sokuhlolwa ukuze unciphise ukucekelwa phansi kwe-RNA okubangelwa ukwethulwa kwe-RNase.

    4. Ama-reagents angcoliswe yi-RNase ngesikhathi sokusetshenziswa.

    Okunconyiwe: Faka esikhundleni Sekhithi Yokuhlukanisa Yezilwane Ephelele Ye-RNA ukuze uthole ukuhlolwa okuhlobene.

    5. Amashubhu e-centrifuge, amathiphu, njll. asetshenziswa ekukhohlisweni kwe-RNA angcoliswe yi-RNase.

    Okutuswayo: Qinisekisa ukuthi amashubhu e-centrifuge, amathiphu, amapayipi, njll. asetshenziswa ekukhishweni kwe-RNA konke Akuna-RNAse.

    I-RNA etholiwe ehlanziwe ithinta ukuhlola okwehla nomfula

    I-RNA ehlanzwe ikholomu yokuhlanza, uma ama-ion kasawoti, okuqukethwe kwamaprotheni kukukhulu kakhulu kuzothinta isilingo esisezansi nomfula, njengokuthi: ukuloba okuhlanekezelwe,Northern Blot et al.

    1. I-RNA ekhishiwe inezinsalela ze-ion kasawoti.

    Okunconyiwe: Qinisekisa ukuthi ivolumu elungile ye-ethanol yengezwe ku-Buffer RW2 futhi wenze amakholomu okuhlanza ama-2 ageza ngesivinini esimaphakathi esikhonjiswe ukusebenza;uma kukhona noma iyiphi insalela ye-ion kasawoti, shiya ikholomu yokuhlanza ku-Buffer RW2 imizuzu engu-5 ekamelweni lokushisa futhi wenze i-centrifugation ukwandisa ukukhishwa kokungcoliswa kukasawoti.

    2. Izinsalela ze-Ethanol ku-RNA ekhishwe.

    Okutuswayo: Qinisekisa ukuthi ngemva kokugeza i-buffer RW2, yenza umsebenzi ongenalutho we-tube centrifugation ngesivinini se-centrifugation esikhonjiswe ukusebenza, ukwandisa isikhathi sokusebenza kwe-tube centrifugation engenalutho sibe yimizuzu emi-2 uma kusekhona insalela ye-ethanol, noma uyishiye ekamelweni lokushisa 5. m

    Bhala umlayezo wakho lapha futhi usithumelele wona