How to calculate the gas composition for a heat treatment where a very low oxygen partial pressure is needed?

I'm interested in a selective oxidation in an austenitic stainess steel so that Chromia rather then ferrous oxide is formed on top of my metal. I know I should have a value of pO2 around...

16 December 2020 8,541 7 View

Why do protein and mRNA levels not always correlate with each other?

Hello, I have performed WB analysis and qPCR analysis to analyze the protein level and the rispective gene relative expression of mTOR, GBA1, LC3 genes in brain of mutant mice. I observed a...

01 December 2020 4,486 2 View

Factor analysis with idependent qualitative variables and dependent quantitative variables ?

Hello, I have a set of independent qualitative variables related to households (type of house, type of windows, presence of pets, ...), and dependent quantitative variables (number of arthropods...

19 April 2020 1,788 11 View

Why does LC3-II not increase after CQ treatment in vivo?

I have administered CQ treatment (50 mg/kg) in twelve-months-old C57BL/6J mice to assess autophagic flux. I validated this protocol on 3-months-old mice first, obtaining an accumulation of LC3II...

05 April 2020 5,018 3 View

What could be the advantages of annealing BaTiO3 in vacuum?

I am evaluating if annealing barium titanate in vacuum could have any benefits.

27 January 2020 4,329 1 View

Cas9 containing plasmid toxic for bacteria?

Hello everyone, I am struggling to obtain an overnight/culture for E.coli Stbl3 bacteria transformed with a plasmid containing the Cas9 enzyme and a GFP donor. Although the bacteria grow well in...

19 January 2020 2,245 3 View

Why lentiviral transduction doesn't work?

Hi everybody. I have a problem trying to overexpress by plasmid the gene of my interest (ffar4 for GPR120). I'm able to infect T293 (they appear totally green), then i filtered and centrifuged the...

28 July 2019 5,831 3 View

Experience with service for gene expression with microarray ?

I am looking for a company which can provide a service for several microarrays for gene expression and subsequent results analyses. Does anyone have any experience to share? E.g. Recommending or...

25 March 2019 1,015 3 View

Heat killing of Bordetella species?

13 March 2018 6,065 2 View

How simulate the behavior of a reinforced concrete beam with a B21 element type?

Hi everyone, I am modelling a reinforced beam under impact load. I chose to model it with element type B21. Any suggestion about how to define material's property in order to simulate...

31 January 2018 7,908 4 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

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

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

Does RNAiMAX transfection reagent work in serum free medium?

I diluted siRNA and RNAiMAX in opti-MEM and added to the cells which they were in the growth medium. Is it a right way? or should I culture cells in the opti-MEM medium for a while and not in...

26 February 2021 10,041 3 View

How do I transfect a large plasmid into PC12 cells with nerve growth factor (NGF) receptor?

I have a large 16 kb plasmid, which I need to transfect into PC12 cells. Lipofectamin 2000 didn't work and with GFP alone the transfection rate is very low. I also tried the neon invitrogen...

25 February 2021 6,635 5 View

What is a good reagent for lentiviral plasmid transfection in HeLa cells?

I have been using Lipofectamine 2000 but was hoping there may be a better protocol or one for something like Lipofectamine PLUS reagent.

24 February 2021 1,633 4 View

Why the cells die after stable transfection?

I tried stable transfection in the U87 cell line with the sgRNA-Cas9 All-in-One lentivector. Cells were cultured in a 6-well plate, 4ug plasmid was transfected after 24hs when the cell confluence...

22 February 2021 2,864 4 View

Is there a method for evaluating nanoparticle adhesion to epithelial tissue in vitro?

I want to measure how well nanoparticles can transfect cells based on NP adhesion properties. In the literature I find people do it in animal studies, and epithelial adhesion helps in transfection...

21 February 2021 7,791 3 View

Are exogenous molecules transferred between human cells during doubling ?

Hello RG community, I wonderer whether exogenous molecules (e.g. reporter plasmids, miRNAs or other artificial molecules) are transferred between cultured human cells during their doubling ? So,...

21 February 2021 6,633 3 View

Which transfection reagent is the best for plasmid transfection of MDA-MB-231 cell line, Lipofectamine based or other reagent suggestion?

I try to transfect MDA MB 231 cell line with a plasmid by PEI, unfortunately, the rate of transfection is low, and now I wanna find and order the best transfection reagent for this cell line and...

19 February 2021 828 6 View