Theoretical computer virology ? is still alive ?

HI Everyone! I am asking to know by you, expert in cybersecurity and mathematics, if Computer virology (the one of Cohen and Andler) is still an active field of research. That research made in...

08 February 2021 7,613 6 View

In opensees is the ReinforcingSteel material sensitive to units of measure?

20 January 2021 7,043 2 View

How do I organize items to repeated surveys for quantitative and qualitative analysis?

Hello fellow researchers! Newbie here :) I created a survey with open-ended and Likert-type questions. The survey was administered nine times to the same people using google forms. I plan to...

01 January 2021 3,921 3 View

Is there a better library or function for plotting CFA with many observed variables in R?

Hello! I'm kind of new in using R for doing EFA, CFA, and SEM. I just followed a guide to perform CFA on my data to see how my context fits the ITC-SOPI model in a VR photo-based tour. I used...

23 December 2020 1,679 2 View

Best way to validate CRISPR-Cas9 editing efficiency?

I'm trying to obtain genetic KO in my cell line of interest using Lentiviral-mediated CRISPR-Cas9 delivery system. I would like to test more than one unpubblished sgRNA targetting a couple of...

10 August 2020 1,324 4 View

How to convert .txt into plink files (ped/map or bed/bim/fam)?

I have a huge oncoarray_genotype_XXXX.txt file with 11.7 GB size. I would like to convert this file into adequate files for analyzing in plink software. However, I could not find how to conduct...

08 July 2020 9,302 2 View

Fluorescence labeled antibody?

Hi all, I have a small question for you. Is there any fluorescent labeled antibody available which emits the fluorescence only if bound to substrate? So the unlabeled version does not emit any...

19 March 2020 5,715 2 View

How to evaporate Methyl Acetate, and purge added Ethanol?

To use an inhibitor, prepared and sold in a methyl acetate solution, I need to evaporate the aforementioned solvent and exchanging to ethanol or DMSO. I will do that with a gentle nitrogen stream....

01 March 2020 6,947 3 View

SPSS, data with 5 replicates. How to generate the variables-data sheet?

I am trying to analyze some results obtained from lipidomics. The experimental setup was as follows: 5 replicates, 3 conditions (1 control, 2 treatments). I have thus 5 numerical values per each...

08 January 2020 3,526 3 View

How to solve multicollinearity problem in mixture design?

Due to its own nature mixture design models (mainly with interactions) presents multicollinearity problems, some authors declare that a combination of forward stepwise and step backward stepwise...

05 January 2020 1,136 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

How to make Fast Green FCF from powder?

Hi everyone We need Fast Green to electroporate plasmid into mice embryos, therefore we simply need it to see some color in the liquid we are electroporating so that electroporation is more...

07 February 2021 1,671 2 View

Does gene insertion via CRISPR/Cas9 result in chromosomal abnormalities?

Hello, I hope to insert a gene into mammalian cells in culture using CRISPR/Cas9 and an HDR template. Will insertion of a 1 kilobase gene cause chromosomal abnormalities and effect cell...

31 January 2021 2,516 1 View

How can I transform 50 kb plasmid into ecoli?

I must put 50 kb plasmid into ecoli strain. I think 50 kb is too big for heat shock with ecoli. Is that correct? Will electroporation work? Or do I need to use cos sites and lambda phage...

22 January 2021 8,612 3 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 the freezing reduces the efficacy of the cells for the electroporation ? should i decrease the voltage for the next time?

I'm making reprogramming of MEFs experiment and I used Piggy bac, Sleeping beauty and episomal transposition systems. After electroporation i froze the cells, then i thawed the cells but there...

06 January 2021 5,467 2 View

Is e.coli ok at Electroporation at 30C?

For reference: I am transforming a newly assembled plasmid what happened: I accidentally incubated my electroporated e.coli cells on the shaker at 30C for 1 hr rather than 37C and did not...

02 December 2020 7,133 5 View

What is the best dye to use to visualise electroporation of neurons with DNA for later expression of a genetically-encoded fluorescent reporter?

We would like to electroporate neurons in organotypic slice cultures with DNA for genetically-encoded flourescent reporters (pH reporters, GEVIs etc). We would like to use an anionic dye that can...

17 November 2020 2,794 1 View

How can I simulate electroporation for a bilayer?

hi I am trying to simulate electroporation for a bilayer containing 10% cholesterol on GROMACS. I have read lots of articles but the only parameter that they have talked about was the electric...

15 November 2020 7,602 2 View

Can I do RNA electroporation without cleanup after in vitro transcription?

I am working on an EMCV like virus with about 8kbp genome. The RNA is usually prepared from digested plasmids by in-vitro transcription, followed by DNase I digestion, followed by P/C and...

02 November 2020 442 3 View