Has someone experience using Lipofectamine Stem Transfection Reagent for transfection of human induced pulripotent stem cells (iPSC)?

07 July 2018 0 2K Report

Badges
Science method

More Andreas Brodehl's questions See All

Who can hep me to analyze WES data for runs of homozygosity?

Hello, we have performed a WES of a single human sample and I am interested to analyse the runs of homozygosity in this sample. Especially, I am interested to unterstand if segmental uniparental...

10 November 2018 7,539 2 View

Does T7 Endonuclease need overlapping nucleotides?

We want to know if Cas9 in combination with a sgRNA was working and have performed a PCR in combination with a T7 Assay. Unfortunately, we could not detect any cleavage product for several sgRNAs....

11 December 2017 2,576 1 View

Which bioinformatic prediction tool could be used to predict the influence of indel mutations?

Hi everyone, which other prediction tools (in addition to Provean) could be used to predict the Impact of indel mutations? Thanks for your help! Andreas

06 July 2017 5,699 3 View

Is it possible to compare one sample with a control set of samples?

Hi everyone, I have measured the RNA of a single patient with a specific rare mutation using qRT-PCR and I have compared it with a control set of probands. My problem is, that I have only one...

04 May 2017 7,264 2 View

What are the best fluorescence proteins for structured illumination microscopy?

Hello everyone, I want to do SIM with three different proteins in living cells. Therefore, I need three different fluorescence proteins with high photostability and high brightness? Could anyone...

02 March 2017 3,617 6 View

What is the definition of stable cell line?

Hi everyone, if I transfect a plasmid with a resistence gene (puromycin) via Lipofectamin (but not with a virus) into eukaryotic cells and select these cells with puromycin over some weeks is the...

02 March 2017 5,470 0 View

Is it possible to use two fluorescence protein tags to increase the brigthness?

Hello everyone, I want to do SIM with living cells and therefore I need fluorescence proteins with high brightness and high photostability. Therefore, I want to ask if someone has fused his/her...

02 March 2017 9,793 3 View

Experience with Bmt1 (restriction enzyme)?

Has anyone experiences with the retriction enzyme BmtI? Are there important specific points to know about this restriction enzyme?

09 October 2014 2,821 1 View

Is it necessary to put the QBI SP163 Enhancer directly upstream of the ATG?

I want to increase the expression of my proteins by inserting the QBI SP163 Enhancer (pcDNA4-Max System, Invitrogen) into another vector system. I want to know if I can add a restriction site (via...

04 May 2014 3,062 0 View

Can you answer my general questions about BiFC?

I am planing BiFC assays and before I want to start I have some general questions: What is the strongest BiFC system? At the moment I could not say anything about the strength of my...

04 May 2014 4,080 3 View

To perform transfection with DharmaFECT Duo in AGS cells. Could you tell me what the ideal concentration is to avoid significant cytotoxicity?

I would like to perform transfection with the reagent DharmaFECT Duo (Horizon) on the AGS cell line. Could you please inform me of the optimal concentration to use without causing cytotoxicity in...

03 August 2024 3,851 1 View

Transfection in HEK293T cells?

Dear All, I am trying to transfect a pCDNA3.1 vector containing my gene of interest. The purpose is to figure out the localization of the protein of interest. I have fused the protein with GFP on...

31 July 2024 9,892 4 View

Issues with Protein Expression in Transfected cells?

Hello I am trying to create a stable cell line in HEK293 via Lipofectamine 3000 transfection. My plasmid is a CD63-IL10-GFP construct with Puromycin resistance. I am successful with the...

30 July 2024 6,648 1 View

How to retain the a GFP tagged gene expression in stable cell line?

Hello , I established a stable cell line expressing GFP tagged to a centrosomal gene having G418 drug selection marker. I validated the stable line by IFA and Western blotting, results are fine....

29 July 2024 5,007 0 View

Does anyone have experience electroporating SKOV3 cells with a large plasmid using Neon transfection?

I have been trying to electroporate SKOV3 cells with a large plasmid (11kb) without much success. Any tips?

29 July 2024 3,229 1 View

Why cannot i find my protein on cell surface after antibiotic selection of expressing plasmid?

I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...

28 July 2024 3,178 3 View

Can I use Polyjet after its expiration date?

I have a Polyjet that has passed its labeled expiration date for 1 year and I'm going to re-start transfection experiments. It is really needed to change it? Transfection efficiency may be lower?

23 July 2024 8,059 1 View

How to transfect circular mRNA to T cells?

Hello! We are using LNP to deliver circular mRNA into T cells. However, compared with linear mRNA, the transfection efficiency and protein expression duration are similar. Are LNP generation or...

21 July 2024 4,322 1 View

Transfection of A549 using lipofectamine 3000?

Im trying to transfect A549 cells using lipfectamine 3000. im using a 96 well plate with 1microgram plasmid DNA in each well . should i remove the culture media or change it or decrease it befor...

21 July 2024 7,610 2 View

Why there is a absence of colonies on the Abx plate for selection after liposome mediated DNA transfer ?

Hi All I am looking to get your opinions, I have perfomed liposome mediated DNA transfer into bacteria, the DNA is a plasmid with msfGFP and when I induce the plasmid I see the fluorescence, and...

16 July 2024 4,698 4 View