Share a copy of RaFD Dataset?

I tried to get this dataset from the original link but no one approved my requests for weeks. Can someone share with me a private link to use? You can send it to my institutional email:...

14 February 2021 1,908 1 View

Help with Amyloid beta detection protocol (western-blotting)?

Hi everyone, I am a first year PhD student working on amyloidosis. I would like some tips and tricks to crack efficiently my current nightmare, i.e. the western blotting of the Amyloid beta and...

11 February 2021 2,513 3 View

Does viral RNA extraction from nasopharyngeal swabs require pre-centrifugation with supernatant collection?

06 February 2021 323 3 View

Aspen plus (hydrometallurgy set-up) & reactions solid-liquid ?

I have to simulate a hydrometallurgical process where there are the following phases: - leaching (solid-liquid reactions) - cementation (solid-liquid reactions) - electrowinning (energy ) -...

30 January 2021 9,225 3 View

How to prediction non-visible parts of microscopic pictures?

Dear all, I like to ask if someone has experience with evaluating pictures towards the prediction of non-visible parts. I have confocal-laser-scanning microscopic pictures of yeast cells and want...

28 December 2020 4,309 11 View

Coronal or Sagittal mouse brain sections to visualise vessels?

Hello, I am a first year PhD student and I am working on Cerebral amyloid angiopathy (CAA). I would like to investigate the presence of my proteins of interest in a CAA/Alzheimer's disease mouse...

05 December 2020 907 3 View

What is the cleanest / best way to handle different software dependencies / configurations?

Dear RG users, I noticed that several toolboxes developed for python are compatible with some specific releases and not with others and in general there is a precise set of tools defining the...

30 November 2020 4,545 5 View

Why maximum allowable limits for toxic metals or heavy metals in agricultural soils vary among countries and institutions?

For example, the maximum allowable limit for USEPA in agricultural soils are very low when compared to Tanzania's maximum allowable limits see figure below. Does this can affect the management...

23 November 2020 5,052 10 View

What is the best way to get organoids of the same size?

I would like to get organoids of the same size. We filter organoids using a 100 um filter and then pass that filtrate through a 40 um filter followed by flipping the 40 um filter. Then we wash the...

22 November 2020 9,391 3 View

How to match Matplotlib color mapping in a R ggscatter plot (or ggplot)?

I created this shinyapp and am having a hard time seeing the image I thought I was...

13 October 2020 223 2 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

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

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

Why is the efficiency of viral mRNA electroporation in PBMC low?

Hello, I have been struggling with the electroporation of PBMC with HIV-I mRNA. Although the quality of the RNA generated is good, and the handling of the cells in meticulous, the frequency of...

27 February 2021 9,233 3 View

Will a 10xHis-tag prevent anti-MBP from binding to MBP?

I have a protein with a 10xHis-MBP tag on the N-terminus. I'm fairly certain I am expressing my protein, but I'm not getting a signal on my western blot. Is there a possibility that the 10xHis tag...

25 February 2021 8,404 3 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

Problem with ligating 2.8 kb insert into 8.5 kb plasmid?

I am trying to clone 2 copies of the same mammalian gene into the pSF-CMV-Ub-Puro Ascl (contains HindIII, KpnI and NheI cut sites) plasmid. This is what the final product is supposed to look like...

24 February 2021 6,310 3 View