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 7,445 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 2,477 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 5,616 3 View

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

04 May 2017 7,165 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 3,522 6 View

What is the definition of stable cell line?

02 March 2017 5,366 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 9,698 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 2,679 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 2,963 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 3,947 3 View

Why might my GFP tag be dissociating from my plasmid after transfection?

I have a set of stably transfected cell lines all transfected with plasmids containing GFP tags on the C terminus. During a western blot using anti-GFP antibody, one of my plasmids has dissociated...

01 March 2021 9,310 4 View

Can someone suggest a way to prepare slides to observe under confocal microscope for observing fluorescent tagged proteins??

I am growing the cells on coverslips and transfecting the fluorescent tagged protein directly on them. Then i want to observe these cells under confocal microscope. Currently i am just using...

01 March 2021 6,142 3 View

Problem with shRNA transfection. Why are only two plates getting contaminated instead of all?

I transfected my LNCaP-WT cells with 3 shRNA plus their NTC two weeks ago and split two puromycin selected cell plates on Friday last week(Feb 26). I checked for GFP in the cells, and they all...

28 February 2021 4,949 3 View

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why the cells die after stable transfection?

I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...

22 February 2021 2,864 4 View

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 3 View

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

Which transfection reagent is the best for plasmid transfection of MDA-MB-231 cell line, Lipofectamine based or other reagent suggestion?

I try to transfect MDA MB 231 cell line with a plasmid by PEI, unfortunately, the rate of transfection is low, and now I wanna find and order the best transfection reagent for this cell line and...

19 February 2021 828 6 View