Lnc-RT Heroᵀᴹ I (ya na gDNase) (Super Premix maka njikọ cDNA nke mbụ sitere na lncRNA)
Nkọwapụta
25×20μl Rxns, 50×20μl Rxns
Lnc-RT HeroTM I (ya na gDNase) bụ usoro ntụgharị ntụgharị ewepụtara nke ọma maka lncRNA iji wepụ nje DNA ngwa ngwa.Ngwakọta 5 × gDNase nwere ike wepu genome fọdụrụ na RNA ngwa ngwa na 42°C maka nkeji 2, na-ezere ndabichi nke genome na nsonaazụ qPCR.
5 × L-RT HeroTM Mix nwere Foregene LncRNA Reverse Transcriptase nke Foregene mebere, nke bụ ụdị ntụgharị ntụgharị ọhụrụ maka lncRNA na ndebiri RNA ndị ọzọ dị ogologo, yana njikọ RNA siri ike yana arụmọrụ ngbanwe dị elu.Sistemu emeziri nke ọma na-eme ka ntụgharị ntụgharị ọsọ ọsọ ma nwee ike ịdegharị ndebiri RNA ngwa ngwa nwere ọdịnaya GC dị elu yana nhazi nke abụọ siri ike.Enwere ike mezue njikọ cDNA nke mbụ na 42 Celsius n'ime nkeji iri na ise.
Ngwa ngwa
| 5× Ngwakọta gDNase |
| 5× Ngwakọta L-RT HeroTM |
| ddH2O na-efu RNase |
| Ntuziaka |
Atụmatụ & uru
■Ikike dị mma iwepu gDNA, nke nwere ike wepu gDNA na ndebiri n'ime nkeji 2.
∎ Ọ nwere ike weghachi ntụgharị nke lncRNA nke ọma.Mgbe ngwaahịa ntụgharị ntụgharị ka edobere na qPCR, uru Ct dị ala karịa nke sistemụ ntụgharị ntụgharị.
∎ Sistemụ ntụgharị ntụgharị nke ọma, ọ na-ewe naanị nkeji iri na ise iji mechaa njikọ nke eriri cDNA mbụ.
∎ ndebiri mgbagwoju anya: ndebiri nwere ọdịnaya GC dị elu yana usoro nke abụọ dị mgbagwoju anya nwekwara ike iji nnukwu arụmọrụ tụgharịa.
∎ Sistemụ ntụgharị ntụgharị uche dị elu, ndebiri ọkwa pg nwekwara ike nweta cDNA dị elu.
■ Sistemụ ntụgharị ntụgharị nwere nkwụsi ike dị elu, ezigbo mmeghachi omume okpomọkụ bụ 42℃, ma ọ ka na-arụ ọrụ ntụgharị ntụgharị dị mma na 50℃.
Ngwa ngwa
Ntụle ọnụọgụgụ nke lncRNA.
lncRNA zuru ezu quantitative analysis.
Ọ nwere ike nyochaa ngwa ngwa na n'ụzọ ziri ezi RNA trace dị ka nje RNA.
Weghachite ndebiri RNA nwere ọdịnaya GC dị elu ma ọ bụ nhazi nke abụọ dị mgbagwoju anya.
Usoro ọrụ
Nchekwa na Shelf ndụ
Ekwesịrị ịchekwa ngwa ahụ na -20°C. Chekwaa ngwaahịa ahụ na refrjiraeto na-ekpo ọkụ mgbe niile na -20°C ozugbo ịnatachara ya.Ọ bụrụ na ọnọdụ nchekwa ahụ dabara adaba, ngwaahịa ahụ agaghị eweda arụmọrụ ọ bụla n'ime oge nkwado afọ 1.
Ntuziaka nyocha nsogbu
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-kpọmkwem nkwalite
1. Nhazi primer enweghị ezi uche
Nkwanye: Chepụta primers dị ka ụkpụrụ nhazi nke mbụ si dị.
2. Ihe fọdụrụ genome.
Aro: Jide n'aka na nzọụkwụ mbụ nke gDNA mmeghachi omume mmeghachi omume okpomọkụ bụ 42ºC, na oge mmeghachi omume nwekwara ike ịgbatị ruo 5min.
Enweghị mgbama nkwalite site na RT-qPCR
1. RNA na-eweda ala.
Nkwanye: Ihe maka mmịpụta RNA kwesịrị ịdị ọhụrụ dịka o kwere mee, yana RNA dị oke mma na ịdị ọcha kwesịrị iji.
2. RNA nwere ihe mgbochi.
Nkwanye: Ndị na-egbochi ntụgharị ntụgharị n'ozuzu gụnyere SDS, guanidine salts, EDTA, wdg. A na-atụ aro ka ịsacha RNA precipitate na 70% ethanol iji wepụ ihe mgbochi.
3. Ihe gbasara imewe nke mbụ.
Nkwanye: Dị ka ụkpụrụ nhazi nke primer si dị, megharịa primers maka nyocha.
Mgbapụta ebumnuche pụtara na njikwa oghere
1. Mmetọ nke ngwaọrụ arụ ọrụ ma ọ bụ reagents.
Nkwanye: Reagents niile ma ọ bụ akụrụngwa maka nnwale kwesịrị ka ejiri akpaaka.Kpachara anya ma dị nro mgbe ị na-ejizi iji gbochie ịmịnye ihe nlele DNA n'ime pipette ma ọ bụ wụfuo na tube centrifuge.
2. Mmetọ mere n'oge nkwadebe nke usoro mmeghachi omume PCR.
Nkwanye: Kpachara anya dị mkpa mgbe ị na-ejizi, dị ka: iyi uwe latex, na-eji ndụmọdụ nzacha.Jiri Ngwakọta PCR Real Time na sistemu na-egbochi mmetọ.
3. The primers na-eweda n'ala.
Aro: Jiri SDS-PAGE electrophoresis chọpụta ma ọ̀tụ̀tụ̀tụ̀ ihe ndị e ji emepụta ihe arụrụala, ma jiri ihe ọ̀tụ̀tụ̀ ọhụrụ dochie anya nnwale nchọpụta fluorescence.
Akwụkwọ ntuziaka:
