...constitutive promoter and can be used to monitor transfection or viral infection efficiency, to study changes... Renilla luciferase is used to monitor viral transfection Feng Zhang 21471 pLenti PGK V5-LUC Neo (w623... Firefly luciferase is used to moniter viral transfection Sidi Chen 109396 TBG-Vector Firefly TBG AAV-... Firefly luciferase is used to moniter viral transfection Sidi Chen 118412 ssAAV-EF1a-FLuc-WPRE-HgHpA_...

...sequencing to confirm library representation. Transfection of the DNA pooled library and appropriate packaging...Addgene.) This small aliquot is not enough to transfect producer cells, so your first step will be to ...in Lenti-X 293T cells using a polyethyenimine transfection protocol. I also recommend searching PubMed ...

...replication. To generate a virus, you need only transfect the ‘helper’ plasmids (viral packaging machinery...Protein production  HEKs are extremely easy to transfect in bulk and produce recombinant proteins at high...

...mammalian selection cassette to screen for positive transfected cells. The addition of an in-frame fluorescent...you can get away with generating transiently transfected cells, you will not need a mammalian selection...

...expression vectors, human embryonic kidney cells are transfected with the plasmid(s) encoding the heavy and light...contain low levels of co-purified plasmid from transfection. This residual plasmid is extracted and amplified... The fluorescence pattern is compared to an untransfected control. Immunohistochemistry (IHC) Tissue sections...antibody. Protein expression is compared to an untransfected control and the protein size is confirmed. Residual...

...constitutively expressed to identify successfully transfected cells. C-to-T editing by the APOBEC-Cas9 editosome... to cells segregated solely by a reporter of transfection, which only report the efficiency of plasmid...frequency of base edits of interest than solely transfected cells. “Gene On” (GO)- a functional reporter ...

...interference (RNAi). The most common method is transfection of short interfering RNA (siRNA). Knockdown .... This approach often involves the transient transfection of expression plasmids in a given cell line....antibody should be able to detect even in cells transfected with low efficacy. But since there can be large...differences between endogenous expression and transfected overexpression, this approach cannot confirm ...

... number of plasmids that are required in the transfection step of the viral packaging cell line. For example...resources on addgene.org Addgene Protocols: General Transfection The Addgene Retrovirus Guide  ...supplied by three types of plasmids, which are cotransfected into the 293T packaging cell line. This system...

...base pairs on each side, in fact. Later, when we transfect the cells, these large regions of homology enable...being cut and destroyed by CRISPR/Cas9 during transfection. Making the double strand break and introducing...

... your packaging cell line, prepped your DNA, transfected and harvested your lentivirus. Now it’s time ...the correct transfer plasmid was used in the transfection; if preparing several different lentiviruses...