The BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System withEmGFP contains either the BLOCK-iT™ Pol II miR RNAi ExpressionVector Kit which contains two boxes. The cloning box containslinearized pcDNA™6.2-GW⁄EmGFP-miR, 10X annealing buffer, T4 DNALigase, 5X DNA Ligation buffer, lacZ control oligo, lacZ controlplasmid, negative control plasmid, DNase/RNase-free water, and forwardand reverse sequencing primers. Store the vectors, buffers, controloligo and plasmids, water, and sequencing primers at -20°C.The One Shot™ box contains transformation reagents includingtwenty-one 50-µl aliquots of One Shot™ TOP10 ChemicallyCompetent E. coli, S.O.C. medium, and a pUC19 supercoiled controlplasmid. Store these transformation reagents -80°C. Allreagents are guaranteed stable for 6 months when properly stored.
The BLOCK-iT™ Lentiviral Pol II miR RNAi Expression System withEmGFP also includes all the necessary reagents for Gateway™ cloninginto the lentiviral expression system, including: pLenti6⁄V5-DESTVector, pLenti6-V5⁄GW-lacZ control, pDONR™221, ViraPower™Lentiviral Bsd Support Kit (ViraPower™ Packaging Mix,Lipofectamine™ 2000 transfection reagent, and Blasticidin), 293FTCell Line, One Shot™ Stbl3™ Chemically Competent E. coli, OneShot™ TOP10 Chemically Competent E. coli, and Gateway™ LR andBP Clonase™ II Enzyme Mixes. Store the Lipofectamine™ 2000reagent at +4°C (do not freeze). Store the vectors,ViraPower™ Packaging Mix, Blasticidin, and both Clonase™ IIEnzyme Mixes at -20°C. Store the competent cells at-80°C. Store the 293FT Cell Line in liquid nitrogen. Allreagents are guaranteed stable for 6 months when properly stored.