Ukuthwala "uMthengi kuqala, umgangatho ophezulu kuqala" engqondweni, sisebenza ngokusondeleyo kunye nabathengi bethu kwaye sibabonelele ngeenkonzo ezisebenzayo kunye namava kwiiMveliso eziNgqongileyo zaseTshayina eziLahlwayo zezoNyango zeNucleic Acid Extraction Reagent DNA/Rna Detection Reagents, iiKits zeNucleic Acid Reagents, Siyakwamkela ukuba usibhalisele omnye komnye ukwenza ishishini lakho libe lula.Ngokuqhelekileyo singamaqabane akho asebenzayo xa ufuna ukuba neshishini lakho.
Qwalasela “uMthengi kuqala, umgangatho ophezulu kuqala” engqondweni, sisebenza ngokusondeleyo nabathengi bethu kwaye sibabonelele ngeenkonzo ezisebenzayo nezinamava kubathengi bethu.China Nucleic Acid Extraction, Indlela ye-Magnetic Bead, Ngoku sineemveliso ezigqwesileyo kunye nezisombululo kunye neentengiso eziqinisekisiweyo kunye neqela lobugcisa.Ngophuhliso lwenkampani yethu, siye sakwazi ukubonelela abathengi ngemveliso engcono kakhulu, inkxaso yobugcisa obuhle, inkonzo egqibeleleyo emva kokuthengisa.

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.

IKholamu ye-RNA kuphela inokubophelela ngokufanelekileyo i-RNA ngefomula ekhethekileyo.Inani elikhulu leisampulu linokucutshungulwa ngaxeshanye.

Amacandelo ekhithi

*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.

Isicelo seKit

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

Ukuhamba komsebenzi

Umzobo

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 ℃ inyanga eyi-1 emva kokufaka i-2-hydroxy-1-ethanethiol (ukhetho). I-Bear "uMthengi kuqala, umgangatho ophezulu kuqala" engqondweni, sisebenzisana ngokusondeleyo nabathengi bethu kwaye sibabonelele ngeenkonzo ezisebenzayo kunye namava kwiiMveliso eziNgqongileyo zaseChina eziLahlwayo zezoNyango zeNucleic Acid Extraction yakho kwi-Accounting entsha ye-Reagent ye-Reagent ye-Reagent ye-Reagent ye-Aclicid i-Reagent entsha nathi omnye komnye ukwenza ishishini lakho libe lula.Ngokuqhelekileyo singamaqabane akho asebenzayo xa ufuna ukuba neshishini lakho.
Iimveliso ezihamba phambiliChina Nucleic Acid Extraction, Indlela ye-Magnetic Bead, Ngoku sineemveliso ezigqwesileyo kunye nezisombululo kunye neentengiso eziqinisekisiweyo kunye neqela lobugcisa.Ngophuhliso lwenkampani yethu, siye sakwazi ukubonelela abathengi ngemveliso engcono kakhulu, inkxaso yobugcisa obuhle, inkonzo egqibeleleyo emva kokuthengisa.

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 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.