I would like to ask some questions about concentrations of Urea 3M and 8M in two experiments?

I would like to ask if anyone knows if concentrations of Urea 3M and 8M can interfere with protein purification at Ni-NTA and Glutathione Sepharose 43 resins. Another question, Urea, at 3M and 8M...

17 November 2020 5,014 3 View

I would like to ask if anyone among you knows what is the ideal concentration of TFA to use at a chemical synthesis of peptides?

During a chemical synthesis of peptides by the F-moc method, the following happened: all the initial steps of the synthesis occurred as planned, but in the cleavage, TFA is used. In the lab, there...

02 November 2020 2,997 8 View

Is there any software to simulate viral infection of cells?

Hello, I am looking for a computational simulator of in vitro infection of cells by virus, to be used for teaching purposes. Do you know any tools/resources ? I would like students to set...

13 April 2020 2,144 1 View

Does anyone has a good experience growing the TALL-104 cell line?

We bought this cell line 6 months ago and we are trying to make it grow, no success so far. We are using ATCC suggested media and replacing IL-2 every 3 days. Fully media changes (by centrifuging...

13 January 2020 7,719 1 View

EO771 mouse tumor transfection protocol?

Hello! I've been trying to transfect EO771 using lipofectamine 2000, but despite changing the transfection media after 6 hours of incubation I have a high rate of cell death. My untransfected...

13 November 2019 7,883 3 View

How can I calculate Ethanol's boiling point for a specific volume?

08 October 2019 7,724 3 View

How can avoid aggregation of cells in E. Coli staining with ThS and propidium iodide when trying to visualize them in an epifluorescence microscope?

I got competent cells of E. Coli induced with IPTG to express B-amyloid peptide and I stained them with ThS and propidium iodide after permeabilizated with Triton x 100; however, when I want to...

07 August 2019 3,158 1 View

No title

I have used FITC-dextran in my mice via gavage, and collected the blood samples to run together. Should I keep the samples at -20C or 4C? If I freeze my FITC can that affect the resulting...

31 July 2019 137 2 View

How quantify relic DNA in water samples?

I need to quantify the amount of relic/free DNA in water samples from a fish pond. I'm looking for a protocol with these method. Does anyone know of any accurate methodology for this...

21 May 2019 5,636 3 View

Which are the best resources to learn AnyLogic?

17 April 2019 7,086 6 View

Which cell line should we use to perform biological dosimetry experiments?

We are preparing some experiments based on irradiating cells under different conditions in order to evaluate the effects in terms of DNA damage, genetic expression, etc. As our project is...

01 March 2021 3,355 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

Precipitate after adding laemmli buffer to protein solution containing Potassium Acetate?

Hi, I am running a size exclusion chromatography experiment with a buffer containing Potassium Acetate as a salt. I analyse these fractions through SDS-PAGE. After boiling my SEC fractions in...

01 March 2021 2,622 3 View

What is the most suitable server for predicting protein structures in bacteria?

I have used the i-Tasser several times, however it has been unavailable for several days. I tried the swiss-model, but the output was not very pleasant due to the model used. Is there any other...

28 February 2021 4,521 3 View

Do you need the CMV enhancer before the CMV promotor for a proper function of CMV on the gene that follows?

I'm a student and I have to produce a cell line with a knockin for the NRF2 gene with GFP. I have to put a promotor in front of the GFP because the gene will be too far away from the promotor of...

28 February 2021 7,127 2 View

Are protein samples in SDS compatible with ELISA?

I have used an AllPrep DNA/RNA/Protein Mini Kit QIAGEN kit to extract RNA and protein from my samples. At the end of the protein purification, I resuspend my protein in 5% SDS. Will these samples...

28 February 2021 7,370 3 View

What is the ideal efficiency for a qPCR?

hello everyone, I need to do standard curves for my qPCR, what is the ideal efficiency range? I tried a primer (Mglu2 receptor) that gave an efficiency of 90.2%. Is it accepted?

28 February 2021 1,254 3 View

Unknown debris sheets observed in cell culture flasks?

Hi, We culture neural progenitor cells in T175 culture flasks coated with PLO-Laminin. We have successfully grown these cells with our protocol for months. Recently, we observed large sheets...

25 February 2021 3,443 4 View

How to replicate the well conditions of a crystal hit with 20%(w/v) isopropanol?

I had a crystal hit in a well with 0.1M HEPEs pH 7.5, 10%(w/v) PEG 4000, and 20%(w/v) isopropanol. Why is isopropanol in w/v here if it's just solvent? How do I recreate this well condition?

25 February 2021 9,761 1 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