MakeyourtargetspecificgRNAlentivectorswithdesiredMarkers!

pLenti-gRNA-U6/H1- CloningKitisdesignedfor cloningofdouble-strandDNAencodingyourdesired gRNAsequenceforCRISPRgenomicgeneediting.Oncetranscribed,thetargetspecificgRNAwillguildCAS9enzymetocutthegenomicDNAatdesiredloci.

TomakeyourtargetspecificgRNAlentivector,twosyntheticgRNAoligonucleotidesare firstannealedtoformthedoublestandDNAduplexwhichis thenclonedintotheprovided,linearpLenti-gRNA-U6 /H1lentivectorviaT4enzymeligation.

ProductFeatures:

1)Flexibility toselectdesiredpromoter: H1orU6;

2)Flexibilitytochoose desired selectionmarker:antibioticorantibiotic-fluorescentfusiondualmarker;

3)Easyandhighefficiencycloningreaction:withprovidedpre-cut,readytouse,lineargRNAlentivector,youdoNOTneedthetedious,yetoftentroublesomelentivectorbackbonepreparation.TheCloningpositiveratecloseto100%.

4)Everythingisincludedinthiskit,enoughforgenerating10gRNAlentivectors;

5)OptionallyusedastetracyclineinducIBLe gRNAtranscription(forH1promoteronly);

ThisproductisthelentivectorversionhavingthegRNAdrivenbyaconstitutivehumanU6 promoter witha GFP-Blastidicin dualselectionmarkerunderRsvpromoter.PleaseseeProductManualfordetails.  

Kitcontents:

pLenti-U6-(GFP-Bsd)linearvector:                 10ul(10rxn,1ul/rxn)
10xgRNAoligoannealingsolution:         25ul
10Xligationbuffer:                      20ul
T4ligaseenzyme:                      10ul(10rxn,1ul/rxn)
Cloningcontrolinsert:annealedNeg-gRNAduplex: 10 ul(10rxn,1ul/rxn)
Sequencingprimer:                                          10ul(10xn,1ul/rxn)

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

ThegeneratedgRNAlentivectorcanbeusedfortransfectionorelectroporationbasedCRISPRediting,orusedforgRNAlentivirusproductionandthenusethegRNAlentivirusfor CRISPRediting. 

Cat#: gRNA-U6-GB