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

TheexpressionofshRNAwasdrivenby anoptionalinduced humanH1promoter.ThisKitprovidesmaterialsfor generating10of yourownshRNAexpressionlentivectors withthefollowingadvancedfeatures. Pleaseseeproductmanualfordetails.

shRNAvectorcloningscheme:    

Kitcontents:

pEco-Lenti-H1-(RFP-Bsd)linearvector                   10ul(10rxn,1ul/rxn)
10xshRNAoligoannealingsolution:                       50ul
5Xligationbuffer:                                                50ul
T4ligaseenzyme:                                                 10ul(10rxn,1ul/rxn)
Cloningcontrolinsert:annealedLuc-shRNAduplex:   5ul(2rxn,1ul/rxn)
Sh-Sequencingprimer:                                           10ul(10rxn,1ul/rxn)

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

KeyFeatures:

1. Linearizedvectorandreadyforuse,noneedforthetediousbenchworksforpreparationofvectorbackbone;