I-Lnc-RT Heroᵀᴹ I(Nge-gDNase)(I-Super Premix ye-cDNA ye-first-strand synthesis esuka kwi-lncRNA)
Iinkcukacha
25×20μl Rxns, 50×20μl Rxns
I-Lnc-RT Hero TM I (Nge-gDNase) yinkqubo yokushicilela eguqulelwe ngokukodwa i-lncRNA ukususa ngokukhawuleza ukungcoliswa kwe-genomic DNA.I-5 × gDNase Mix inokususa ngokukhawuleza i-genome eseleyo kwi-RNA kwi-42 ° C kwimizuzu ye-2, igweme ngokufanelekileyo ukuphazamiseka kwe-genome kwiziphumo ze-qPCR.
I-5 × L-RT Hero TM Mix iqulethe i-Foregene LncRNA Reverse Transcriptase ephuhliswe ngokukodwa yi-Foregene, luhlobo olutsha lwe-reverse transcriptase ngokukhethekileyo kwi-lncRNA kunye nezinye iitemplates ezide ze-RNA eziyinkimbinkimbi, kunye nobudlelwane obunamandla be-RNA kunye nempumelelo ephezulu yokurekhoda.Inkqubo ephuculweyo yenza ireyiti yokukhuphela umva ngokukhawuleza kwaye inokukhuphela ngokulula iitemplates zeRNA ezinomxholo ophezulu weGC kunye nesakhiwo sesibini esintsonkothileyo.I-strand yokuqala ye-cDNA synthesis ingagqitywa kwi-42 ° C ngemizuzu eyi-15.
Amacandelo ekhithi
| 5× Umxube we-gDNase |
| 5× I-L-RT Hero TM Mix |
| RNase-Free ddH2O |
| Imiyalelo |
Iimpawu&izinto eziluncedo
■Ikhono elisebenzayo lokususa i-gDNA, enokususa i-gDNA kwi-template ngaphakathi kwemizuzu emi-2.
■ Iyakwazi ukubuyisela umva ngokufanelekileyo ushicilelo lwe-lncRNA.Xa imveliso ekhutshelweyo ebuyela umva iphantsi kwe-qPCR, ixabiso le-Ct lisezantsi kunelo lesiqhelo lenkqubo yokukhuphela umva.
■ Inkqubo esebenzayo yokukhuphela umva, kuthatha imizuzu eli-15 kuphela ukugqiba udityaniso lwe-cDNA yomtya wokuqala.
■ Iithempleyithi ezintsonkothileyo: iithempleyithi ezinomxholo ophezulu weGC kunye nolwakhiwo lwesibini oluntsonkothileyo lunokuguqulwa kwakhona ngobuchule obuphezulu.
■ I-high-sensitivity reverse transcription system, i-pg-level templates inokufumana i-cDNA ekumgangatho ophezulu.
■ I-reverse transcription system inozinzo oluphezulu lwe-thermal, elona qondo lobushushu lokusabela liyi-42℃, kwaye isenomsebenzi olungileyo wokukhutshelwa umva kwi-50℃.
Isicelo seKit
Uhlalutyo lobungakanani be-lncRNA.
lncRNA uhlalutyo olupheleleyo lobungakanani.
Inokukhawuleza nangokuchanekileyo ukuhlalutya umkhondo we-RNA njengentsholongwane ye-RNA.
Reverse transcription ye RNA templates with high GC content or complex complex structure.
Ukuhamba komsebenzi
Ugcino kunye nobomi beShelf
Ikhithi kufuneka igcinwe kwi -20°C. Gcina imveliso kwifriji yokushisa rhoqo kwi -20°C ngoko nangoko emva kokufunyanwa.Ukuba iimeko zokugcina zifanelekile, imveliso ayiyi kuthoba nayiphi na intsebenzo ngexesha le-1 leminyaka yokuqinisekisa.
Isikhokelo sokuHlalutya ingxaki
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-ulwandiso oluthile
1. Uyilo lweprimer olungenangqiqo
Isincomo: Yila iiprimers ngokwemigaqo yoyilo lweprimer.
2. Iintsalela zeGenome.
Ingcebiso: Qinisekisa ukuba inyathelo lokuqala lobushushu bokususwa kwe-gDNA yi-42 ° C, kwaye ixesha lokuphendula linokwandiswa ukuya kwi-5min.
Akukho phawu lokukhulisa nge-RT-qPCR
1. I-RNA ithotyiwe.
Ingcebiso: Izinto zokukhutshwa kwe-RNA kufuneka zibe zintsha kangangoko, kwaye kufuneka kusetyenziswe i-RNA ekumgangatho ophezulu kunye nococeko oluphezulu.
2. I-RNA iqulethe ii-inhibitors.
Isincomo: I-reverse transcription inhibitors ngokubanzi ibandakanya i-SDS, i-guanidine salts, i-EDTA, njl. Kucetyiswa ukuba uhlambe i-RNA precipitate nge-70% ye-ethanol ukususa i-inhibitors.
3. Imiba yoyilo lokuqala.
Isincomo: Ngokomgaqo woyilo lweprimer, yila ngokutsha iiprimers ukuze zihlolwe.
Ibhendi ekujoliswe kuyo ibonakala kulawulo olungenanto
1. Ukungcoliswa kwezixhobo zokusebenza okanye ii-reagents.
Isincomo: Zonke ii-reagents okanye izixhobo zovavanyo kufuneka zifakwe kwi-autoclaved.Qaphela kwaye uthambe xa uphethe ukuthintela iisampulu ze-DNA ukuba zifunxe kwi-pipette okanye zichitheke ngaphandle kwetyhubhu ye-centrifuge.
2. Ungcoliseko lwenzekile ngexesha lokulungiselela inkqubo yokusabela kwe-PCR.
Isincomo: Thatha izilumkiso eziyimfuneko xa uphethe, njengoku: ukunxiba iiglavu zelatex, usebenzisa iingcebiso zokucoca.Sebenzisa Ixesha langempela le-PCR Mix kwinkqubo yobungqina bokungcola.
3. Iiprimers zithotyiwe.
Iingcebiso: Sebenzisa i-SDS-PAGE i-electrophoresis ukubona ukuba iiprimers zonakalisiwe, kwaye endaweni yazo iiprimer ezintsha zovavanyo lokubona i-fluorescence.
Incwadi yoMyalelo:
