Expression cloning by using cDNA,do I need to modify the cDNA first?

I am going to have a expression cloning of mammalian gene by using shuttle plasmid to transforming the E.coli However I don't know I should only inserting the Coding sequence ,or I can...

28 February 2021 5,440 3 View

Schrodinger Desmond runs fine on one GPU, but crashes on other GPU?

Hi, We are having issues using Desmond on our workstation. Simulations run fine on one of our gpus ("Gpu0") but crash consistently on the other gpu ("gpu1" - which is also set up as the display...

01 February 2021 5,866 3 View

How does climate change affect birds (shorebirds)?

I m recently investigating the impact of climate change on shorebird population trends, I would like to know what is the impact of fluctuations in variables such as sea surface temperature, global...

10 November 2020 9,293 15 View

Can I use the EPIC-Oxford FFQ for free?

Designing my PhD project and need to use an FFQ, but we don't have any funding for this part of the project.

09 November 2020 4,541 1 View

Is there a survey tool (similar to REDCap) through which we can send an anonymous survey and restrict respondents to one survey each?

I am working on a research survey where I need to send it out to a group of users and track in (anonymous) responses. I need to restrict users to completing the survey only once. I do not think...

06 August 2020 918 2 View

Western Blot Transfer Band Smearing?

Anyone have any insight? It's becoming a recurring issue. I make fresh transfer buffer each time.

29 July 2020 9,331 9 View

How many users can i put in the same vsat broadband plan?

sorry if i wasn't able to explain very well for example, i am in a country that doesn't have satellite internet service, and would like to be the first provider for this service. lets say i have a...

05 July 2020 4,637 1 View

Failed gateway cloning?

Hi everyone, I am having a few doubts about gateway cloning. We are using gateway cloning for protein expression using pDEST15 as our destination vector. I attached a picture of the pDEST15...

10 March 2020 7,472 2 View

Inrush control when Transformers are Energized at Solar Renewable is best achieved how?

Increasing use of solar renewable resources or farms on weak remote locations are causing significant power quality and voltage notch issues with connecting utilities. High penetration of these...

17 August 2019 7,097 2 View

Glass microballoons from sodium silicate solution?

I once produced microballoons from sodium silicate solution (water glass). does anyone have experience in manufacturing these, are articles you could point me in the direction of? thanks

01 July 2019 8,764 1 View

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03 March 2021 5,360 2 View

Does PHA-P stimulate the proliferation of lymphocytes from mice?

Hello, I will start performing MLR using lymphocytes from mice's spleen and I would like to know the better way for the stimulation. If is using PHA-P or CD3/CD28? I would like to know as well if...

23 February 2021 5,476 4 View

Looking for advices and tips to improve the staining and the gating strategy in human blood?

Hi, I am staining a human blood with CD19, CD3,CD14 and CD16. My gating strategy in FlowJo goes like this: - First, I gate a general population SSC-A VS FSC-A => Then gate for single cells, =>...

21 February 2021 5,724 4 View

How T cells recognise non-peptide antigens?

This might sound very basic question, but please share with me an explanation of how the T cell is activated when the antigen is not a peptide? What happens? And what are the molecules involved in...

21 February 2021 564 3 View

FDC and FRC cell populations in mouse spleen?

I was trying to isolate fibroblastic reticular cells (FRC) and follicular dendritic cells from mice spleen, but kept getting poor yield. I found from literature that the frequency of FDC is around...

17 February 2021 3,506 4 View

How do I avoid artifacts during paraformaldehyde fixation of mouse CD8+ T cells?

I work with mouse OT-I CD8+ T cells which have been activated in vitro with peptide-MHC and B7 proteins for 3 days. I've been trying to standardize protocols to fix these cells using...

28 January 2021 8,519 2 View

Does re-stimulation of T cells with PMA and Ionomycine before staining for intracellular cytokines for flow cytometry affect reliability of the assay?

I understand that to be able to measure intracellular cytokines there is a need to restimulate with PMA and Ionomycin but I don't understand how the results will be reliable after that. PMA and...

18 January 2021 7,240 7 View

Can someone help me identify the bands?

The samples are replicates (4) of total RNA from HEK-293 T cells. The ladder is a 1kBp ladder on a 1% native agarose gel buffered with 1x TAE. From my searches, there should be two sharp bands at...

15 January 2021 4,712 4 View

Does anyone have problems performing CRISPR KO in human primary T cells?

For the past few months, I have been troubleshooting a CRISPR KO of human CD3e in human primary T cells. I got the KO to work one time using 1250 ng TrueCut Cas9 (Thermo) + 250 ng (or 7.5 pmol)...

11 January 2021 6,016 3 View

Do murine T-cells get transient activation when they attach to a tissue culture treated plate?

I have been trying to activate CD19 CAR-T cells using plate-bound anti-CAR Abs. I am getting activation both in cell lines and primary CD8 T-cells. I am using pErk as a readout for activation. I...

30 December 2020 1,655 2 View