How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Can I project all raw motor imagery EEG data onto two spatial filters developed from training data?

Greetings RG community, I have been working on a pipeline for the classification of EEG motor imagery signals. This is currently being done on the Giga Science MI dataset with 52 subjects, using...

28 January 2021 6,947 2 View

How to calculate error for a Log2 ratio of mean freqeuncies?

Hello al! I am looking to calculate and plot error (SD, SEM, etc) for comparing the change in frequency for many immune cell populations between TumorType1 vs TumorType2 (indolent breast tumor vs...

29 September 2020 8,332 1 View

How do I interpret an interaction between variables in a linear mixed model?

For example, I have a 2-level factor variable (diagnose) in interaction with a 6-level factor variable (layer). Intercept by default is the first level of all. In my case intercept is control...

18 September 2020 6,211 4 View

Is there a functional difference between a two-way ANOVA and a one-way ANOVA followed by Tukey's post-hoc?

Hi, I have been thinking about this for a couple months, and I am wondering if anyone else has any input. Lets say I have 4 treatments 1) 0 nM compound X + 0 nM Compound Y 2) 10 nM compound X + 0...

18 August 2020 8,583 5 View

Why do scRNAseq pipelines (e.g. seurat) have the filtering step to remove cells that express over 2500 genes? Meaning, why 2500?

Does anyone know how this number became "default"? what is it based on? How deep is our knowledge of "average" , "healthy" gene expression of different cell types? Sorry if my question is trivial...

04 August 2020 6,691 1 View

Has anyone had success with antibody conjugation kits for ICC/IHC studies?

I am planning a multiplex ICC experiment. The issue is that several of my antibodies are anti-rabbit which would cross react with secondary antibodies. I was wondering if anyone has used antibody...

23 March 2020 6,136 3 View

Is there another site which acts as a forum for device process engineers to discuss recipes/approaches/observations/tips/etc.?

I am well aware that recipes we may come across in a journal article or elsewhere rarely work properly in a different location with a different tool, however, it shows that a thing is possible and...

26 February 2020 2,827 5 View

How can I run and interpret a mixed effect model with a random factor in Stata?

Hi all, I have a set of data (attached) for which I need to run a mixed model with a group variable as a random factor. The data are grouped by the column titled "Group Variable". The group...

13 January 2020 9,398 5 View

Is anyone able to source a canadian either TLR 4 KO, TLR 2 KO, MyD88 KO, or IRF3 KO cell lines?

Hi, I am looking for immune cells with any of the above knocked out, before I look into a CRISPR system to do it myself. I have only been able to source mice with these genes knocked out, but I...

17 November 2019 1,957 2 View

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02 March 2021 3,060 3 View

Issues with Permeabilization during Flow Cytometry?

Hello Everyone. Currently I am working to characterize macrophages in the myocardium after ischemia-reperfusion injury in rats. Due to the low total cell number isolated from rat hearts I can...

01 March 2021 3,867 3 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

Is it possible for two human neuronal cell lines to have different gene sequence for the same protein?

I am looking at the ATP1A2 (Sodium/Potassium ATPase alpha subunit 2) in two human neuronal cell lines. Expression levels of this protein seems to be almost equal when detected by one antibody....

01 March 2021 3,607 3 View

Is CD44 degraded in lysosome together with hyaluronan after CD44 mediated endocytosis of hyaluronic acid?

Also when RHAMM binds hyaluronic acid, they get internalized, will RHAMM also be degraded? Or both CD44 and RHAMM will be transferred back to the cell membrane? Asking for breast cancer cell line...

01 March 2021 8,169 2 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

What could be some reasons why unstimulated monocytes are getting activated?

Im doing PBMC isolation -> CD14+ enrichment using magnetic beads -> stimulation setup. My negative control is just cells in cRPMI but they seem to get activated over and over again.

28 February 2021 7,883 3 View

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28 February 2021 9,936 2 View

Has anyone had luck using cell lines other than HEK 293 (such as COS1) to package lentivirus?

I am looking to package lentivirus in endothelial cells but I am opposed to using 293 cells due to personal ethics regarding the source. I have done some reading and saw that COS1, COS7, & CHO...

25 February 2021 3,030 1 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