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

How vaccines of covid 19 work?

Millions ask how vaccines of covid 19 be successful and safe, its here to explain generally how vaccine of pfizer work ,mRNA vaccines teach our cells how to make a protein that will trigger an...

28 February 2021 2,723 4 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

RIA. How to measure?

When you use RIA, with a control the unknown sample with antibodies, you bare in mind the binding sites of the antibodies. However you still need to measure labelled antigen (radioactive) and not...

28 February 2021 2,133 3 View

What is the most suitable resuspension buffer for TCA/acetone precipitated protein?

I need to extract protein from fermented carbon sources for Bradford assay. Most researchers experiencing insolubility of pellet in resuspension buffer. Please assist me to select most suitable...

26 February 2021 7,129 3 View

EMSA troubleshooting. Protein-DNA complex not migrating, cold probe lane with multiple bands. Can anyone help?

I'm running an EMSA to show the DNA binding activity of recombinant proteins versus the Wild type. Previous assays run by my research group using a different test system show the protein-DNA...

24 February 2021 8,325 1 View

Best plasmid could be used as a standard curve for AAV titration?

I have been trying to have a good AAV titration for my production and I have tried different plasmids to set up as a standard curve but each one behaves differently. i am trying to use ITR for...

23 February 2021 5,167 3 View

How to control the variants vof covid 19 in second and third waves?

A the world find means to control covid 19 by vaccination ,new variants came to spread and make the world in fear and danger, to understand the conduct of these variants, es constantly change...

21 February 2021 3,080 6 View

Surprisingly low antibody titers against SASRS-COV-2 RBD in hamsters compared to mice?

21 February 2021 6,958 2 View

Will SARS-CoV-2 Spike be broken into S1 and S2 by denaturing conditions?

Hello, I am performing a western blot of my SARS-CoV-2 Spike-pseudotyped lentiviral particles. I denature my virus by adding concentrated virus to 2X Laemmli buffer at 1:1, then boiling for...

17 February 2021 4,844 4 View

Does anyone know, can i do cryosectioning of whole aorta after oil red O staining and use the cryosection parts for further analysis?

I would like to quantify the lesions on aorta. I want to take image of atherosclerotic plaques formation in aorta by using O red Oil Solution. After the staining, what will be the storage...

16 February 2021 5,876 2 View