The introduction of DNA into a recipient eukaryote cell and its subsequent integration into the recipient cells chromosomal DNA. | Contact experts in Transfection to get answers
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Questions related to Transfection
I transfected FAM-labelled negative control siRNA (Bioneer, SN-1021) into J774A.1 murine monocyte cells to check the transfection efficiency. I used RNAiMAX (Thermo) 0.6 ul per well of 96-well...
22 December 2021 2,961 1 View
Dear all, I am attaching pictures of microfluidic culture of MCF7 on day 2, day 3 and day 5. I found on day 2 the spheroids had tight boundary but the next day all of a sudden the spheroids are...
20 December 2021 6,451 0 View
I have generated iPSC colonies from peripheral blood, today is Day14 post-transduction and the colonies seem to have acquired nice borders and distinct iPSC morphology (images attached). There is...
17 December 2021 7,058 1 View
Dear everyone, I want to label THP-1 cells for IF microscopy but cannot find any detailed protocol on how to label suspension cells without cytospin - is there at all a way of labelling...
14 December 2021 5,269 3 View
Question related to stable transfection of cells using puromycin
13 December 2021 806 3 View
Hello everyone I have a problem with my transfection protocol in hippocampal neurons between 5 to 7 DIV. I need to transfect a GFP construct. I am currently using the lipofectamine 3000...
13 December 2021 7,298 1 View
Hello, Does anyone used TetR-based Tet-on expression system in E18 rat hippocampal neurons? I want to transfect the hippocampal neurons at 0 d.i.v and treat them with doxycycline at 1 d.i.v. Does...
13 December 2021 9,459 1 View
What is the shelf life of glutamine in the media with glutamin mixed already? And does it have any impact on cells after being denatured?
13 December 2021 1,997 1 View
I have liquid recombinant protein. What is the best formulation for recombinant protein stability during freezing?
11 December 2021 8,447 6 View
Dear community, In my bachelor thesis I am looking into 4 channel IF stainings of the mouse hippocampus. Until now I had to manually count the marker positive cells in the stacks using Fijis cell...
11 December 2021 1,403 4 View
I am Yasuhiro Endo , a medical doctor in Japan. I am going to incubate HUVEC with 10 ng/ml tumor necrosis factor-α (TNF-α; R&D systems, Abingdon, UK) or PBS as control for 5hrs and measure...
07 December 2021 4,105 0 View
Hi everyone, I just wonder that I combined the DNA mixture with Lipofectamine in the Eppendorf tube, then I stand it at room temperature for 20-30 min (as protocol) in the clean bench of cell...
04 December 2021 8,907 5 View
I have the whole genome of the cultivars from the Phaseolus genus, some chromosomes have annotations about the genes that appear in those chromosomes, while others do not have those. In some...
03 December 2021 7,285 2 View
Hi everyone! These vacuoles was frequently seen in recently cultured U-2OS cells, which greatly affected the imaging results. What are those? How should I do to avoid these vacuoles? U-2OS was...
03 December 2021 1,341 0 View
Hi, I used for the first time the Annexin V-FITC Apoptosis Detection Kit (BioVision) but I am a bit unsure of the results. I investigated the apoptosis on mussel M. edulis haemocytes, by using...
03 December 2021 4,426 3 View
I used the Flp-InTM T-RExTM System to stably transfect 293T cells so that my gene of interest is inducibly expressed by tetracycline. I want to transiently transfect with a luciferase reporter...
02 December 2021 4,517 1 View
Hello, I am working with gut microbes that are quite difficult to lyse. By using a kit, it gives a very low yield. So I am trying out my protocol for total RNA extraction. The extracted RNA will...
02 December 2021 5,404 2 View
I've been doing co-IP to determine which proteins interact with a German cockroach protein known as Bla g 2. However, after preforming LC-MS on the co-immunoprecipitated proteins, I've noticed...
01 December 2021 1,630 3 View
I would want to perform a Mass spectrometric analysis of co-immunoprecipitated proteins after having pulled down a specific protein using magnetic beads (dyna beads) or agarose conjugated with...
01 December 2021 1,866 0 View
These dots are extremely small and do not move, however upon DAPI staining, I observe a lot of DAPI spots surrounding the nucleus and cells. Can it be mycoplasma contamination? I do not observe...
30 November 2021 6,157 5 View
Hi, Can anyone share their transfection protocol for NIH 3T3 cells. I tried with lipo3000 protocol. Very few cells are transfected and their shapes change after transfection.
30 November 2021 6,582 1 View
As mentioned in the title, how to maintain cells in hypoxia conditions? The special incubator is not always available. Furthermore, I would like to keep cobalt chloride away, a commonly used...
28 November 2021 5,722 3 View
Hello! I have a question regarding my AAV experiments. I tried to transduce my HEK293T cells with AAV8. I already tested my vector with a gRNA in vitro and had a good knock-down rate, while also...
26 November 2021 5,020 3 View
Hello Scientists, I am working on assaying the half life of a plasma membranal protein which is transiently transfected into Hek293T cell line using FuGene-HD transfection reagent. Kindly, if...
25 November 2021 2,327 1 View