I-Lnc-RT Heroᵀᴹ I(With gDNase)(Super Premix for first-strand cDNA synthesis from lncRNA)
Imininingwane
25×20μl, 50×20μl rxn
I-Lnc-RT HeroTM I (Nge-gDNase) iwuhlelo lokuloba oluhlanekezelwe ngokukhethekile ukuze i-lncRNA isuse ngokushesha ukungcoliswa kwe-genomic DNA.I-5×gDNase Mix ingasusa ngokushesha i-genome esele ku-RNA ku-42°C imizuzu engu-2, igweme ngempumelelo ukuphazamiseka kwegenome emiphumeleni ye-qPCR.
I-5×L-RT HeroTM Mix iqukethe i-Foregene LncRNA Reverse Transcriptase ethuthukiswe ngokukhethekile i-Foregene, okuwuhlobo olusha lwe-reverse transcriptase eqondiswe ngokukhethekile i-lncRNA nezinye izifanekiso eziyinkimbinkimbi ze-RNA ezinde, ezinokuhlobana okuqinile kwe-RNA nokusebenza kahle kokurekhoda okuhlehlayo okuphezulu.Isistimu ethuthukisiwe yenza isilinganiso sokuhlehla sokuloba sisheshe futhi ingabhala kalula izifanekiso ze-RNA ezinokuqukethwe okuphezulu kwe-GC kanye nesakhiwo sesibili esiyinkimbinkimbi.I-strand cDNA synthesis yokuqala ingaqedwa ku-42 ° C ngemizuzu eyi-15.
Izingxenye zekhithi
| 5× I-gDNase Mix |
| 5× I-L-RT HeroTM Mix |
| RNase-Free ddH2O |
| Iziyalezo |
Izici&izinzuzo
■Ikhono elisebenzayo lokususa i-gDNA, engasusa i-gDNA kusifanekiso phakathi nemizuzu emi-2.
■ Ingakwazi ukuhlehlisa ngempumelelo ukulotshwa kwe-lncRNA.Uma umkhiqizo wokulotshwa okuhlanekezelwe ungaphansi kwe-qPCR, inani le-Ct liphansi kunalelo lesistimu yokubhala ngokuhlehla evamile.
■ Isistimu yokubhala ehlehlayo esebenza kahle, kuthatha imizuzu eyi-15 kuphela ukuqedela ukuhlanganiswa kwe-cDNA yomucu wokuqala.
■ Izifanekiso eziyinkimbinkimbi: izifanekiso ezinokuqukethwe okuphezulu kwe-GC nesakhiwo sesibili esiyinkimbinkimbi nazo zingahlehliswa ngendlela esebenza kahle kakhulu.
■ Isistimu yokubhala ehlehlayo ezwelayo ephezulu, izifanekiso zezinga le-pg nazo zingathola i-cDNA yekhwalithi ephezulu.
■ Isistimu yokubhala ngokuhlehlayo inokuqina okuphezulu kwe-thermal, izinga lokushisa elilungile lokusabela lingu-42℃, futhi isenokusebenza okuhle kokulotshiweyo okuhlehla kokungu-50℃.
Isicelo sekhithi
Ukuhlaziywa komthamo okuhlobene kwe-lncRNA.
I-lncRNA i-absolute quantitative analysis.
Ingakwazi ukuhlaziya ngokushesha nangokunembile ukulandelela i-RNA njengegciwane le-RNA.
Ukuhlehlisa ukulotshwa kwezifanekiso ze-RNA ezinokuqukethwe kwe-GC ephezulu noma ukwakheka kwesibili okuyinkimbinkimbi.
Ukugeleza komsebenzi
Isitoreji kanye Nempilo yeshelufu
Ikhithi kufanele igcinwe ku -20°C. Gcina umkhiqizo esiqandisini sokushisa esingashintshi ku -20°C ngokushesha ngemva kokuthola.Uma izimo zesitoreji zifanelekile, umkhiqizo ngeke wehlise noma yikuphi ukusebenza phakathi nesikhathi sokuqinisekisa sonyaka ongu-1.
Umhlahlandlela Wokuhlaziya Inkinga
The following is an analysis of the problems that may be encountered in the use of Lnc-RT HeroTM I (With gDNase) series kits, hoping to be helpful to your experiments. In addition, we have dedicated technical support to help you with other experimental or technical problems beyond the operating instructions and questions. If you have any needs, please contact us: 028-83361257 or E-mail: Tech@foregene.com.
Non-i-amplification ethize
1. Idizayini ye-primer engenangqondo
Okunconyiwe: Dizayina ama-primers ngokuya ngemithetho yokuklama yokuqala.
2. Izinsalela ze-genome.
Ukusikisela: Qiniseka ukuthi isinyathelo sokuqala se-gDNA yokushisa yokusabela ekususeni ngu-42°C, futhi isikhathi sokusabela singanwetshwa sibe yi-5min.
Ayikho isignali yokukhulisa nge-RT-qPCR
1. I-RNA yehlisiwe.
Okutuswayo: Impahla yokukhipha i-RNA kufanele ibe yintsha ngangokunokwenzeka, futhi i-RNA yekhwalithi ephezulu nehlanzeke kakhulu kufanele isetshenziswe.
2. I-RNA iqukethe ama-inhibitors.
Isincomo: Ama-reverse transcription inhibitors ngokuvamile ahlanganisa i-SDS, usawoti we-guanidine, i-EDTA, njll. Kunconywa ukugeza i-RNA precipitate nge-ethanol engu-70% ukuze kukhishwe ama-inhibitors.
3. Izinkinga ze-Primer design.
Okunconyiwe: Ngokuhambisana nesimiso sokuklama kokuqala, hlela kabusha ama-primer ukuze ahlolwe.
Ibhendi eqondiwe ivela kusilawuli esingenalutho
1. Ukungcoliswa kwamathuluzi okusebenza noma ama-reagents.
Okunconyiwe: Wonke ama-reagents noma okokusebenza kokuhlolwa kufanele kufakwe ngokuzenzakalelayo.Qaphela futhi ube mnene lapho uphatha ukuze uvimbele amasampula e-DNA ukuthi angamuncwa ku-pipette noma achitheke ngaphandle kweshubhu le-centrifuge.
2. Ukungcola kwenzeke ngesikhathi sokulungiswa kohlelo lokusabela kwe-PCR.
Okutuswayo: Thatha izinyathelo zokuphepha ezidingekayo lapho uphatha, njengokuthi: ukugqoka amagilavu e-latex, usebenzisa amathiphu okuhlunga.Sebenzisa I-Real Time PCR Mix kusistimu yokuvikela ukungcola.
3. Iziqalo zehlisiwe.
Isiphakamiso: Sebenzisa i-SDS-PAGE electrophoresis ukuze uthole ukuthi iziqalisi zonakalisiwe, futhi esikhundleni salokho ufake iziqalo ezintsha zokuhlolwa kokutholwa kwe-fluorescence.
Incwadi Yeziqondiso:
