pEco-Lenti-H1-shRNA-(Puroomycin)CloningKitisdesignedtocloningofdouble-strandDNAencodingashorthairpinRNA.Oncetranscribed,theshRNAwasprocessedintoshortRNAinvivoforRNAianalysis.TomakeshRNAexpressingvector,twosyntheticoligonucleotideswerefirstannealedtoformthedoublestandDNAduplexwhichwasthenclonedintotheReady-to-use,linearpEco-Lenti-H1-(Puromycin)vectorviaT4enzymeligation.

ThetranscriptionofshRNAwasdrivenbyamodifiedhumanH1promoter,aRNApolymeraseIIIpromoter.ThemodifiedH1promoterisanoptionalinducIBLepromoterwhichcaneither constitutivelyexpressshRNAorinduciblyexpressshRNAbytetracyclineinduction.

ThisKitprovidesmaterialsforgenerating10 ofyourownshRNAexpressionlentivectorwiththefollowingadvancedfeatures. Pleaseseeproductmanualfordetails.

shRNAvectorcloningscheme:   

Kitcontents:

pEco-Lenti-H1-(Puromycin)linearvector:              10ul(10rxn,1ul/rxn)
10xshRNAoligoannealingsolution:                       50ul
5Xligationbuffer:                                                 50ul
T4ligaseenzyme:                                                 10ul(10rxn,1ul/rxn)
Cloningcontrolinsert:annealedLuc-shRNAduplex:   5ul(5rxn,1ul/rxn)
Sh-Sequencingprimer:                                          10ul(10rxn,1ul/rxn)

Materialrequiredbutnotincluded:
DH5achemicalcompetentcells:                        10vials(10xn,1vial/rxn)

KeyFeatures:

1. Linearizedvectorandreadyforuse,noneedforthetediousbenchworksforpreparationofvectorbackbone;