Skip to main content

Questions tagged [transfection]

The tag has no usage guidance.

Filter by
Sorted by
Tagged with
3 votes
1 answer
4k views

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 ...
rhill45's user avatar
  • 3,705
2 votes
1 answer
248 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 ...
The Nightman's user avatar
  • 1,836
2 votes
0 answers
48 views

How many copies of RNA per cell are usually reached through overexpression in human cell-lines? (any technique)

After 8 hours of online-research I was unable to find any info at all.. I was able to get some concrete copy numbers of DNA (e.g. plasmid) per cell after transfection of diverse transfection ...
KaPy3141's user avatar
  • 1,597
2 votes
0 answers
757 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 ...
Marta Tesin's user avatar
2 votes
0 answers
597 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 ...
molecular.me's user avatar
2 votes
0 answers
397 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 ...
user137's user avatar
  • 5,348
1 vote
2 answers
513 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 ...
P M's user avatar
  • 19
1 vote
1 answer
291 views

In plasmid-based transient transfection of mesenchymal stem cells, do I have to select for transfected cells and verify GOI expression via a reporter?

I am transiently transfecting mesenchymal stem cells with a mammalian plasmid-based expression vector that does not contain any mammalian selection marker and also the gene of interest will be cloned ...
Kane Yane's user avatar
0 votes
2 answers
89 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?...
M Rahman's user avatar
0 votes
0 answers
40 views

Why does hemagglutinin transfection fail in HEK293 cells?

I have tried to transfect HEK293 cells with the Influenza A H1N1 PR8 Hemagglutinin expression plasmid. https://www.sinobiological.com/cdna-clone/h1n1-hemagglutinin-ha-vg11684-ch I used PEI MAX as the ...
Beginner's user avatar