Questions tagged [transfection]

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Cationic lipid mediated transfection optimization for 150mm dishes

I need to optimize a transfection protocol to transiently express a plasmid encoding a chimera of eyfp attached to the c term of a Golgi apparatus signaling molecule) in hela cells and hepg2 cells ...
2
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1answer
228 views

Retrovirus Production

I have been having difficulties with low transduction efficiencies of my retrovirus production. I expand my plasmid of interest (on MiG-GFP plasmid) in DH5α E Coli for ~24 hours, purify with Qiagen ...
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0answers
437 views

What's the best wat to combine biological replicates in qPCR gene expression analysis? (2^(-∆∆Ct) method; one-way anova, turkey's test)

I am measuring the levels of gene x after transfection of cells with a plasmid and I want to compare them to the levels in mock-transfected cells. I did three different transfection experiments, in ...
2
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0answers
533 views

Scaling down amount of DNA for transfection: what factor to consider?

I am interested in transfecting HEK293 cells with a BAC. One paper suggests using 24 ug of DNA to transfect a 55-cm^2/10 ml culture plate. I am using an assay optimimzed for a 96 well plate so I am ...
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2answers
341 views

293T safety, transfections with plasmid DNA

I am a little concerned with 293T cell lines, I have been working with this cell line for a few years with less frequency. However there were times that i extracted the proteins from these cell lines ...
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0answers
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final control prior to transfecting an expression vector into a mammalian cell?

I have been asked a general question: Once I have cloned a full-length cDNA into an expression vector, what final important control must I do before I transfect this into an embryonic stem cell line? ...
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0answers
374 views

Transfect Mammalian Cells with Single Stranded DNA?

Agrobacteria can deliver parts of their DNA to plant cells as T-DNA, transfer DNA. This DNA is delivered as a single strand and can be integrated into the plant genome or can be converted to a double ...
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2answers
83 views

Luciferase promoter vector over p-AcGFP1-C1 vector

AcGFP1 vector can emit light by itself, whereas in case of Luc vector a substrate is needed for the reaction. Nevertheless, Luc is said to be more specific or better than AcGFP1. Why is this so called?...