pEco-Lenti-U6-shRNA-(Blasticidin)CloningKitisdesignedtocloningofdouble-strandDNAencodingashorthairpinRNA.Oncetranscribed,theshRNAwasprocessedintoshortRNAinvivoforRNAianalysis.TomakeshRNAexpressingvector,twosyntheticoligonucleotideswerefirstannealedtoformthedoublestandDNAduplexwhichwasthenclonedintotheReady-to-use,linearpEco-Lenti-U6-(Bsd)vectorviaT4enzymeligation.

ThetranscriptionofshRNAwasdrivenby aconsitutivehumanU6promoter.ThisKitprovidesmaterialsforgenerating10ofyourownshRNAexpressionlentivectors withthefollowingadvancedfeatures. Pleaseseeproductmanualfordetails.

shRNAvectorcloningscheme:    

Kitcontents:

pEco-Lenti-U6-(Blastidicin)linearvector:              10ul(10rxn,1ul/rxn)
10xshRNAoligoannealingsolution:                        50ul
5Xligationbuffer:                                                50ul
T4ligaseenzyme:                                                 10 ul(10rxn,1ul/rxn)
Cloningcontrolinsert:annealedLuc-shRNAduplex:  5ul(5rxn,1ul/rxn)
Sh-Sequencingprimer:                                          10ul(10rxn,1ul/rxn)

Materialrequiredbutnotincluded:
DH5aorotherchemicalcompetentcells:                  10vials(10xn,1vial/rxn)

KeyFeatures:

1. Linearizedvectorandreadyforuse,noneedforthetediousbenchworksforpreparationofvectorbackbone;