Received plasmid pVP-EL222 from University of Texas Southwestern

pVP-EL222 resuspended (sample from University of Texas Southwestern)

pVP-EL222 transformed into competent cells

pVP-EL222 did 3 in 1

pVP-EL222 extracted from transformed cultures and checked with restriction digestion (EcoRI andXhoI)

pVP-EL222 plasmid stored and transformed cultures cryopreserved

pVP-EL222 amplified with plasmid PCR (contaminated NC)

pVP-EL222 checked with plasmid PCR with brand new reagents (success, no contamination detected)

pVP-EL222 amplified with plasmid PCR

pVP-EL222 digested with SpeI and BamHI)

pVP-EL222 amplified with PCR

pVP-EL222 amplified with PCR than digested

pVP-EL222 checked separately with plasmid PCR with redesigned primers

pVP-EL222 checked with plasmid PCR due to multiple failed ligations

pVP-EL222 checked with restriction digestion due to multiple failed ligations

pVP-EL222 checked with Taq polymerase plasmid PCR with remixed primers

pVP-EL222 checked with KOD polymerase plasmid PCR with remixed primers

pVP-EL222 amplified with PCR (use for ligation with both pBAR_PgpdA and PMCL1)

pVP-EL222 amplified with PCR (use for ligation with both pBAR_PgpdA and PMCL1)

pVP-EL222 amplified with PCR (use for ligation with both pBAR_PgpdA and PMCL1)