Ukuphishekela kwethu kwaphakade isimo sengqondo esithi "bhekisa imakethe, bhekisa isayensi" kanye nokholo oluyisisekelo "ngecala lokuhlobisa okuthuthukile, uma unesisekelo sokuthi uzwakale ngokuphelele mayelana nenhlangano yethu noma nge-imeyili yakho, kufanele ukwaziswa kakhulu.
Esiphishekelayo saphakade isimo sengqondo "sokubheka imakethe, bheka isiko, bheka isayensi" kanye nethiyori "yekhwalithi eyisisekelo, nokholo ekuqaleni kanye nokuphatha okuthuthukile"I-China Nucleic Acid Extraction, Ikhithi yokukhipha i-Rna, Siyazihlonipha njengenkampani ehlanganisa ithimba eliqinile lochwepheshe abasungula izinto ezintsha futhi abanolwazi olunzulu ekuhwebeni kwamazwe ngamazwe, ukuthuthukiswa kwebhizinisi kanye nokuthuthuka kwemikhiqizo.Ngaphezu kwalokho, inkampani ihlala ihlukile phakathi kwezimbangi zayo ngenxa yezinga layo eliphakeme lekhwalithi ekukhiqizeni, kanye nokusebenza kahle kwayo nokuvumelana nezimo ekusekelweni kwebhizinisi.

Kits Izincazelo

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Le kit isebenzisa ikholomu ye-spin kanye nefomula eyakhiwe inkampani yethu, engakwazi ukukhipha i-RNA ephezulu kanye nekhwalithi ephezulu engqikithini yezilwane ezihlukahlukene ngokusebenza kahle okuphezulu.Ihlinzeka ngekholomu ye-DNA-Cleaning ephumelelayo, engakwazi ukuhlukanisa kalula futhi ikhangise i-DNA ye-genomic ku-supernatant kanye ne-tissue lysate, elula futhi elondoloza isikhathi;Ikholomu ye-RNA kuphela ingahlanganisa ngempumelelo i-RNA futhi ingacutshungulwa kanyekanye ngefomula eyingqayizivele Amasampuli amaningi.

Lonke uhlelo luyi-RNase-Free, ukuze i-RNA ekhishiwe ingonakalisiwe;I-Buffer RW1, i-Buffer RW2 isistimu yokugeza ibhafa, ukuze i-RNA etholiwe ingabi nawo amaprotheni, i-DNA, i-ion, kanye nokungcoliswa kwenhlanganisela yezinto eziphilayo.

Izingxenye zekhithi:

Izici&izinzuzo

■ Asikho isidingo sokukhathazeka ngokuwohloka kwe-RNA;lonke uhlelo luyi-RNase-Free
■ Khipha ngempumelelo i-DNA usebenzisa i-DNA-Cleaning Column
■ Khipha i-DNA ngaphandle kokwengeza i-DNase
■ Kulula-yonke imisebenzi iqedwa ekamelweni lokushisa
■ Ukusebenza okusheshayo kungaqedwa ngemizuzu engama-30
■ I-Safe-akukho reagent ephilayo edingekayo
■ Ukuhlanzeka okuphezulu -OD260/280≈1.8-2.1

Isicelo sekhithi

Ilungele ukukhishwa nokuhlanzwa kwe-RNA ephelele ezinhlobonhlobo zezicubu zezilwane ezintsha noma eziqandisiwe noma amaseli akhulisiwe.

Imingcele yomkhiqizo

■ Izinhlelo zokusebenza eziwela phansi: ukuhlanganiswa kwe-cDNA ye-first-strand, i-RT-PCR, i-molecular cloning, i-Northern Blot, njll.
■ Amasampula: izicubu zezilwane, amaseli akhulisiwe
■ Umthamo: Izicubu 10-20mg, Amaseli(2-5)×10⁶
■ Umthamo omkhulu wokubopha we-DNA wekholomu yokuhlanza: 80 μg
■ Ivolumu ye-Elution: 50-200 μl

Ukugeleza komsebenzi

Umdwebo

I-Animal Total RNA Isolation Kit iphathwe ama-20mg
Amasampula egundane amasha, thatha i-RNA ephelele engu-5% 1% ye-agar

I-Glycogel electrophoresis
1: Ubende 2: Izinso
3: Isibindi 4: Inhliziyo

Isitoreji nempilo yeshelufu

Ikhithi ingagcinwa izinyanga ezingama-24 ekamelweni lokushisa (15-25 ℃) noma 2–8 ℃ isikhathi eside.I-Buffer RL1 ingagcinwa ku-4 ℃ inyanga engu-1 ngemva kokwengeza i-β- mercaptoethanol (uyazikhethela).Izinto esiziphishekelayo zaphakade isimo sengqondo "sokubheka imakethe, ukubheka isiko, ukubheka isayensi" kanye nethiyori "yekhwalithi eyisisekelo, yiba nokholo ekuqaleni kanye nokuphatha okuthuthukisiwe" kwe-Ordinary Discount China Nucleic Acid Extraction 96 I-Testnatic Acid DNA I-Testne Magt uma unokubuyekezwa mayelana nenhlangano yethu noma izimpahla, qiniseka ukuthi uzizwe ukhululekile ukukhuluma nathi, i-imeyili yakho ezayo izobongwa kakhulu.
Isaphulelo EsijwayelekileI-China Nucleic Acid Extraction, Ikhithi yokukhipha i-Rna, Siyazihlonipha njengenkampani ehlanganisa ithimba eliqinile lochwepheshe abasungula izinto ezintsha futhi abanolwazi olunzulu ekuhwebeni kwamazwe ngamazwe, ukuthuthukiswa kwebhizinisi kanye nokuthuthuka kwemikhiqizo.Ngaphezu kwalokho, inkampani ihlala ihlukile phakathi kwezimbangi zayo ngenxa yezinga layo eliphakeme lekhwalithi ekukhiqizeni, kanye nokusebenza kahle kwayo nokuvumelana nezimo ekusekelweni kwebhizinisi.

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 ngendlela eyanele nokuthi amaseli ezicubu ahlukaniswe ngokwanele ukuchaza ukukhululwa kwe-RNA.

4. I-eluent ayengezwanga ngendlela efanele.

Okunconyiwe: Qinisekisa ukuthi i-RNase-Free ddH₂U-O wengezwa ngokudonsela phansi maphakathi nolwelwesi lwekholomu 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 kanye ne-Buffer RW2 bese uxuba kahle ngaphambi kokusebenzisa ikhithi.

6. Umthamo wesampula wezicubu 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 ukukhishwa okungaphelele.

Okutuswayo: Ivolumu ye-elution yekholomu yokuhlanza ingu-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.

Okutuswayo: Uma kunensalela ye-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 emi-5 ukuze kukhishwe ngokwanele i-ethanol esele.

I-RNA ehlanziwe 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 ingxenye eyodwa yezingcezu lapho uzisebenzisa ukugwema ukuncibilika okubandayo kwesampula kanye nokonakala kwe-RNA.

3. I-RNase yethulwa noma ingagqoki 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 Ikhithi Yokuhlukanisa Yezilwane Ephelele Yenani Le-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 ehlanjululwe ngekholomu 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 elutioned.

Isincomo: Qinisekisa ukuthi ngemva kokugeza kwe-buffer RW2, yenza i-tube centrifugation operation engenalutho ngesivinini se-centrifugation esiboniswe ukusebenza, ukwandisa isikhathi sokusebenza kwe-tube centrifugation engenalutho kumaminithi angu-2 uma kusekhona insalela ye-ethanol, noma uyishiye ekamelweni lokushisa imizuzu engu-5 ngemva kwe-tube centrifugation engenalutho ukuze ukhulise ukukhishwa kwe-thanol.