IMIQATHANGO YAKHO YOPHUPHA "Sijonge isiko, jonga isayensi" kunye nethiyori ye-China yokuCoca i-Adneid ye-nud ye-nugney ye-nugney ye-nugney ye-nugney ye-nugney ye-nugness i-Innacy "yePosi ye-Special Dead" ye-imeyile iya kuxatyiswa.
Usukelo lwethu lwanaphakade sisimo sengqondo “sokujonga imarike, qwalasela isiko, jonga inzululwazi” kunye nethiyori “yomgangatho wesiseko, yiba nokholo kulawulo lokuqala kunye nolawulo oluphambili”China Nucleic Acid Extraction, Rna Extraction Kit, Siyazizukisa njengenkampani equka iqela elomeleleyo leengcali ezivelisa izinto ezintsha nezinamava kurhwebo lwamazwe ngamazwe, uphuhliso lweshishini kunye nokuphuculwa kwemveliso.Ngaphezu koko, inkampani ihlala iyingqayizivele phakathi kwabakhuphisana nabo ngenxa yomgangatho ophezulu womgangatho kwimveliso, kunye nokusebenza kwayo kunye nokuguquguquka kwinkxaso yoshishino.

Iinkcazelo Kits

50 Preps, 200 Preps

Le kit isebenzisa ikholomu ye-spin kunye nefomula ephuhliswe yinkampani yethu, enokuthi ikhuphe ubunyulu obuphezulu kunye nomgangatho ophezulu we-RNA kwizicubu zezilwanyana ezahlukeneyo kunye nokusebenza kakuhle.Inika iKholamu ye-DNA-Cleaning esebenzayo, ekwazi ukuhlula ngokulula kwaye ibhengeze i-DNA ye-genomic ukusuka kwi-supernatant kunye ne-tissue lysate, elula kunye nokugcina ixesha;IKholamu ye-RNA kuphela inokubophelela ngokufanelekileyo i-RNA kwaye inokucutshungulwa ngaxeshanye ngefomula eyodwa Iisampuli ezininzi.

Yonke inkqubo i-RNase-Free, ukuze i-RNA ekhutshiweyo ingathotywanga;I-Buffer RW1, i-Buffer RW2 inkqubo yokuhlamba i-buffer, ukwenzela ukuba i-RNA efunyenweyo ayinayo iprotheni, i-DNA, i-ion, kunye nongcoliseko lwe-organic compound.

Amacandelo ekhithi:

Iimpawu&izinto eziluncedo

■ Akukho mfuneko yakuzikhathaza ngokuthotywa kweRNA;yonke inkqubo i-RNase-Free
■ Ukususa ngokufanelekileyo i-DNA-usebenzisa i-DNA-Cleaning Column
■ Susa i-DNA ngaphandle kokufaka i-DNase
■ Ilula-yonke imisebenzi igqityezelwa kubushushu begumbi
■ Ukukhawuleza -ukusebenza kunokugqitywa kwimizuzu engama-30
∎ Ekhuselekileyo-akukho zixhobo zokuvuselela izinto eziphilayo ezifunekayo
■ Ukucoceka okuphezulu -OD260/280≈1.8-2.1

Isicelo seKit

Ilungele ukutsalwa kunye nokucocwa kwe-RNA iyonke ukusuka kwiintlobo ezahlukeneyo zezicubu zezilwanyana ezitsha okanye ezikhenkcezisiweyo okanye iiseli ezikhulisiwe.

Iiparamitha zemveliso

■ Izicelo ezisezantsi: i-first-strand cDNA synthesis, RT-PCR, i-molecular cloning, i-Northern Blot, njl.
■ Iisampulu: izihlunu zezilwanyana, iiseli ezikhuliswayo
■ Umthamo: Izicubu 10-20mg, Iiseli(2-5)×10⁶
■ Ubuninzi be-DNA yokubopha umthamo wekholomu yokucoca: 80 μg
■ Umthamo we-Elution: 50-200 μl

Ukuhamba komsebenzi

Umzobo

Animal Total RNA Isolation Kit baphathwa 20mg
Iisampulu ezintsha zemouse, thatha i-5% esulungekileyo iyonke i-RNA 1% ye-agar

Glycogel electrophoresis
1: Udakada 2: Izintso
3: Isibindi 4: Intliziyo

Ukugcinwa kunye nobomi beshelufu

Ikhithi ingagcinwa iinyanga ezingama-24 kwiqondo lobushushu begumbi (15–25 ℃) okanye 2–8 ℃ ixesha elide.I-Buffer RL1 ingagcinwa kwi-4 ℃ kwinyanga ye-1 emva kokongeza i-β- mercaptoethanol (ukhetho) .Izinto zethu ezingunaphakade zizimo zengqondo "ngokubhekiselele kwimarike, ngokubhekiselele kwisiko, ngokubhekiselele kwisayensi" kunye nethiyori "yomgangatho osisiseko, nokholo lokuqala kunye nolawulo oluphambili" lwe-Ordinary Discount China Nucleic Acid Extraction kunye ne-Testnetic Acid 96 ye-Testnetic ye-DNA ye-Kitnetic ityala unalo naluphi na uphononongo malunga nombutho wethu okanye iimpahla, qiniseka ukuba unamava akhululekileyo ukuthetha nathi, i-imeyile yakho ezayo iya kuxatyiswa kakhulu.
Isaphulelo EsiqhelekileyoChina Nucleic Acid Extraction, Rna Extraction Kit, Siyazizukisa njengenkampani equka iqela elomeleleyo leengcali ezivelisa izinto ezintsha nezinamava kurhwebo lwamazwe ngamazwe, uphuhliso lweshishini kunye nokuphuculwa kwemveliso.Ngaphezu koko, inkampani ihlala iyingqayizivele phakathi kwabakhuphisana nabo ngenxa yomgangatho ophezulu womgangatho kwimveliso, kunye nokusebenza kwayo kunye nokuguquguquka kwinkxaso yoshishino.

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 yokusebenza kwe-centrifugation linganyuswa ukuya kwi-2min, okanye ikholomu yokucoca inokubekwa kwindawo yokushisa ye-5 min ukususa ngokwaneleyo i-ethanol eseleyo.

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. Ukunyibilika 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 ayinxibi 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 ye-centrifugation operation kwisantya se-centrifugation esiboniswe ukusebenza, ukwandisa ixesha le-tube engenanto ye-centrifugation operation ukuya kwi-2 min ukuba kusekho i-ethanol intsalela, okanye uyishiye kwindawo yokushisa kwe-5 min emva kwe-tube engenanto ye-centrifugation yokwandisa ukususwa kwe-ethanol ye-ethanol.