Sihlala sicabanga futhi sizijwayeza okuhambisana nokushintsha kwesimo, futhi sikhule.Sihlose ukuzuza ingqondo nomzimba ocebile kanye nokuphila Kwabadayisi Bezitolo Ezithengisa Ngaphakathi BeProfessional Magnetic Viral Rna Nucleic Acid Isolation Purification Prepacked Kits.Sithemba ukuthi singaba nobudlelwano obuhle nosomabhizinisi abavela kuzo zonke izindawo ezizungezile.
Sihlala sicabanga futhi sizijwayeza okuhambisana nokushintsha kwesimo, futhi sikhule.Sihlose ukuzuza ingqondo nomzimba ocebile kanye nokuphilaI-China Universal Rna DNA Reagents kanye ne-Automatic Nucleic Acid Purification, Sifisa ukuhlangabezana nezidingo zamakhasimende ethu emhlabeni jikelele.Uhlu lwethu lwempahla namasevisi lukhula ngokuqhubekayo ukuze luhlangabezane nezidingo zamakhasimende.Samukela amakhasimende amasha namadala avela kuyo yonke imikhakha yempilo ukuthi asithinte ukuze athole ubudlelwano bebhizinisi besikhathi esizayo futhi azuze impumelelo efanayo!
Amamanuwali wemiyalo:

 Sihlala sicabanga futhi sizijwayeza okuhambisana nokushintsha kwesimo, futhi sikhule.Sihlose ukuzuza ingqondo nomzimba ocebile kanye nokuphila Kwabadayisi Bezitolo Ezithengisa Ngaphakathi BeProfessional Magnetic Viral Rna Nucleic Acid Isolation Purification Prepacked Kits.Sithemba ukuthi singaba nobudlelwano obuhle nosomabhizinisi abavela kuzo zonke izindawo ezizungezile.
Abathengisi bezitolo ezinkulu zeI-China Universal Rna DNA Reagents kanye ne-Automatic Nucleic Acid Purification, Sifisa ukuhlangabezana nezidingo zamakhasimende ethu emhlabeni jikelele.Uhlu lwethu lwempahla namasevisi lukhula ngokuqhubekayo ukuze luhlangabezane nezidingo zamakhasimende.Samukela amakhasimende amasha namadala avela kuyo yonke imikhakha yempilo ukuthi asithinte ukuze athole ubudlelwano bebhizinisi besikhathi esizayo futhi azuze impumelelo efanayo!

Umhlahlandlela Wokuhlaziya Inkinga

Ukuhlaziywa okulandelayo kwezinkinga ongahlangabezana nazoIngqikithi YezitshaloRNA extraction will help you with your experiments. In addition, for other experimental or technical problems in addition to operating instructions and problem analysis, we have dedicated technical support to help you. If you have any needs, please contact us at: 028-83360257 or E-mali : Tech@foregene.com.

Ikholomu ejikelezayo ivalekile

Ukuvinjwa kwekholomu yokuphonswa kuzobangela ukuthi isivuno se-RNA sehliswe noma singakwazi ukuhlanzwa ukuze kutholwe i-RNA, futhi ikhwalithi ye-RNA etholiwe izoba phansi.

Ukuhlaziywa kwembangela evamile:

1. Isampula ayiphukile ngokuphelele.

Ukuhlukaniswa kwesampula okungaphelele kungavimba Ikholomu Yokuhlanza I-DNA, engaphinda ithinte isivuno nekhwalithi ye-RNA.Sincoma ukuthi lapho wenza ukuhlukaniswa kwesampula, ugaye ngokushesha inani elanele le-nitrogen ewuketshezi ukuze uhlukanise izicubu ezifana nezindonga zamaseli kanye nolwelwesi lwamaseli lamasampuli ngangokunokwenzeka.Kumasampula ezitshalo e-polyphenol polysaccharides, sincoma ukuthi usebenzise i-Plant Total RNA Isolation Kit Plus.

2.Aspirate the DNA-Cleaning Column isolated supernatant, aspirate kungenzeka cell cell doti pellet.

I-aspirated cell debris pellet ingavala Ikholomu ye-RNA kuphela phakathi nezinqubo ze-RNA adsorption (bona isinyathelo 5 senqubo, isinyathelo 6 senqubo ye-polysaccharide polyphenol).Sincoma ukuthi ukunakekelwa kufanele kuthathwe lapho ufisa lesi sikhulu esinamandla ukuze ugweme ukungcola kwamaseli.

3. Inani lokuqala lesampula likhulu kakhulu.

Ukusetshenziswa kwesampula okweqile kuzoholela ekuhlukaneni kwesampula okungaphelele noma ukuhlaziya okungaphelele kwamaseli nge-Buffer PRL1 noma i-Buffer PSL1, okuholela kukholomu yokuhlanza evalekile yemisebenzi yokuhlanza.I-Plant Total RNA Isolation Kit inomkhawulo wokuqala ongu-50 mg ngokuhlanzwa okukodwa kwesampula esebenzayo.Ngamasampuli ezitshalo ze-polyphenol polysaccharides, sincoma ukuthi uzame i-Plant Total RNA Isolation Kit Plus.

4. Izinga lokushisa le-centrifuge liphansi kakhulu.

Ukuhlukaniswa nokuhlanzwa okuphelele kwe-RNA ngaphandle kokuphazamiseka kwe-nitrogen ewuketshezi yesampula yezicubu, zonke izinyathelo zenziwa kumazinga okushisa asekamelweni (20-25 °C).Amanye ama-centrifuge ezinga lokushisa eliphansi anamazinga okushisa angaphansi kuka-20 °C, angabangela ukuvinjelwa Kukholomu Yokuhlanza I-DNA kanye/noma Ikholomu ye-RNA kuphela.Uma lokhu kwenzeka, setha izinga lokushisa le-centrifuge libe ngu-20-25 °C bese ufudumeza ingxube ye-lysis kanye/noma ukufakwa kwe-ethanol supernatant yokuhlukanisa ibe ngu-37 °C.

Ayikho i-RNA ekhishiwe noma isivuno se-RNA siphansi

Ngokuvamile kuba nezinto eziningi ezithinta ukusebenza kahle kokuthola kabusha, njengalezi: isampula yokuqukethwe kwe-RNA, indlela yokusebenza, ivolumu ye-elution, njll.

Ukuhlaziywa kwezimbangela ezivamile njengoba ngezansi:

I-1.Ukugeza kweqhwa noma izinga lokushisa eliphansi (4 ° C) i-centrifugation yenziwa ngesikhathi sokusebenza.

Ukusikisela: Sebenzisa ekamelweni lokushisa (15-25 ° C) kuyo yonke inqubo, ungawagezi iqhwa kanye ne-centrifugation yokushisa ephansi.

       2.I-RNA yehlisiwe ngenxa yokulondolozwa okungafanele kwesampula noma ukulondolozwa kwesikhathi eside kwesampula.

Okutuswayo: Amasampula asanda kuqoqwa kufanele aqandwe ngokushesha ku-nitrogen ewuketshezi, abese egcinwa ku -80°C isikhathi eside, agweme ukuqhwaza okuphindaphindiwe nokuncibilika kwamasampuli;noma ngokushesha cwilisa amasampula ku-RNA stabilizer RNAlater solution (amasampula ezilwane).

       3.Ukuhlukaniswa kwesampula okunganele kanye ne-lysis kuholela ekuvinjweni kwekholomu yokuhlanza.

Isiphakamiso: Uma ugaya izicubu, sicela uqinisekise ukuthi isicubu sigaywe ngokwanele, futhi usidlulisele ngokushesha ku-Buffer PSL1 esilungiselelwe ngaphambilini (qinisekisa ukuthi ingxenye elungile ye-β-ME yengeziwe, bona isinyathelo 1 senqubo).

4.I-eluent yengezwe ngokungalungile.

Ukusikisela: Qinisekisa ukuthi i-ddH ye-RNase-Free₂U-O uconselwa phakathi nolwelwesi lwekholomu yokuhlanza.

5.Ivolumu elungile ye-ethanol ephelele ayizange yengezwe ku-Buffer PSL2 noma ku-Buffer PRW2.

Ukusikisela: Sicela ulandele imiyalelo, wengeze ivolumu efanele ye-ethanol ephelele ku-Buffer PSL2 kanye ne-Buffer PRW2 bese uxuba kahle ngaphambi kokuba ikhithi isetshenziswe.

6.Inani lesampula lethishu alifanelekile.

Isiphakamiso: Sebenzisa u-50 mg wethishu ngo-500 μl we-Buffer PSL1.Ukusebenzisa izicubu eziningi kuzonciphisa inani le-RNA ekhishiwe futhi ukuhlanzeka kwe-RNA okuwumphumela nakho kuzoncipha.Sincoma ngokuqinile ukuthi umthamo wokuqala wesampula akufanele udlule u-50 mg ngokusebenza kokukhipha kwe-RNA ngakunye.

7. Ivolumu ye-elution engafanele noma ukungezwani okuphelele.

Isiphakamiso: Umthamo ongacacile wekholomu yokuhlanza ungama-50-200 μl;uma umphumela we-elution ungagculisi, kunconywa ukuthi unwebe isikhathi kuzinga lokushisa legumbi ngemva kokwengeza i-ddH eshisiwe ye-RNase-Free₂O, njengemizuzu emi-5-10.

8.Ikholomu yokuhlanza inensalela ye-ethanol ngemva kokugeza nge-Buffer PRW2.

   Isiphakamiso: Uma ishubhu elingenalutho li-centrifuged iminithi elingu-1 futhi kusekhona i-ethanol esele ngemva kokugeza ku-Buffer PRW2, ungakwazi ukwandisa isikhathi se-tube centrifugation engenalutho sibe yimizuzu emi-2, noma ubeke ikholomu yokuhlanza ekamelweni lokushisa imizuzu emi-5 ukuze ususe ngokuphelele i-ethanol esele.

9.Ikhithi isetshenziswe ngokungalungile.

   Isiphakamiso: Kumasampula ezitshalo we-polyphenolic polysaccharides, ukusebenzisa amakhithi avamile njenge-Plant Total RNA Isolation Kit kungase kungakwazi ukuthola amasampula e-RNA afanelekile.Sincoma ukuthi usebenzise i-Plant Total RNA IsolationKit Plus, eklanyelwe ngokukhethekile amasampula ezitshalo ze-polyphenolic polysaccharide.Ikhithi eyakhelwe ngokukhethekile ukukhipha i-RNA ku-polyphenol namasampula ezitshalo ze-polysaccharide.

OD260/OD280 inani liphansi

Ukukhishwa kwe-RNA nge-ddH₂O futhi okusetshenziselwa ukufundwa kwe-spectrophotometer kubangela amanani aphansi e-OD260/OD280.Sincoma ukusebenzisa i-10 mM Tris-HCl, i-pH 7.5 (kune-ddH ye-RNase-Free₂O ukuze uthole i-RNA) ukuze uthole amanani afanele e-OD260/OD280, bheka “I-RNA Concentration and Purification Assays” ekhasini 19.

I-RNA ehlanziwe yehlisiwe

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

Ukuhlaziywa kwezimbangela ezijwayelekile:

1.Amasampula ezicubu awazange agcinwe ngesikhathi ngemuva kokuqoqwa.

    Okutuswayo: Uma amasampula ezicubu engasetshenziswa ngesikhathi ngemva kokuqoqwa, sicela uwagcine ku-nitrogen ewuketshezi ezingeni lokushisa eliphansi ngokushesha noma uwadlulisele ku--80°C ukuze agcinwe isikhathi eside ngemva kokuqhwaza okusheshayo ku-nitrogen ewuketshezi, noma ngokushesha ucwilise amasampula ku-RNA stabilizer RNAlater solution (amasampula ezilwane ).Ukuze uthole i-RNA, zama ukusebenzisa amasampula ezicubu ezisanda kuqoqwa.

2.Ukuqhwaza okuphindaphindiwe nokuncibilika kwamasampula ezicubu.

   Ukusikisela: Lapho ugcina amasampula ezicubu, kungcono ukuwasika abe izingcezu ezincane ukuze alondolozwe, futhi ukhiphe ingxenye yawo lapho uwasebenzisa ukuze ugweme ukuwohloka kwe-RNA okubangelwa ukubanda okuphindaphindiwe nokuncibilika kwamasampula.

3.I-RNase yethulwa egunjini lokuhlinza noma engagqoki amagilavu ​​alahlwayo, imaski, njll.

   Isiphakamiso: Ukuhlolwa kokukhipha i-RNA kwenziwa kangcono kakhulu emisebenzini ehlukene ye-RNA, futhi ithebula laselabhorethri kufanele lihlanzwe ngaphambi kokuhlolwa, futhi amagilavu ​​alahlwayo namamaski kufanele kugqokwe phakathi nokuhlolwa ukuze kugwenywe ukuwohloka kwe-RNA okubangelwa ukwethulwa kwe-RNase ngezinga elikhulu kakhulu.

4.I-reagent ingcoliswe yi-RNase ngesikhathi sokusetshenziswa.

   Isiphakamiso: Faka esikhundleni ngochungechunge olusha lwamakhithi okukhipha i-RNA esitshalo esiphelele sokuhlolwa okuhlobene.

5.Amashubhu e-centrifuge namathiphu we-pipette asetshenziselwa ukukhwabanisa kwe-RNA angcoliswe yi-RNase.

Ukusikisela: Qinisekisa ukuthi amashubhu e-centrifuge, amathiphu e-pipette, ama-pipette, njll. asetshenziswa ekukhishweni kwe-RNA konke Akuna-RNAse.

Amamanuwali wemiyalo:

I-Plant Total RNA Isolation Kit Instruction Manual