I would like to ask some questions about concentrations of Urea 3M and 8M in two experiments?

I would like to ask if anyone knows if concentrations of Urea 3M and 8M can interfere with protein purification at Ni-NTA and Glutathione Sepharose 43 resins. Another question, Urea, at 3M and 8M...

17 November 2020 5,014 3 View

I would like to ask if anyone among you knows what is the ideal concentration of TFA to use at a chemical synthesis of peptides?

During a chemical synthesis of peptides by the F-moc method, the following happened: all the initial steps of the synthesis occurred as planned, but in the cleavage, TFA is used. In the lab, there...

02 November 2020 2,997 8 View

Is there any software to simulate viral infection of cells?

Hello, I am looking for a computational simulator of in vitro infection of cells by virus, to be used for teaching purposes. Do you know any tools/resources ? I would like students to set...

13 April 2020 2,144 1 View

Does anyone has a good experience growing the TALL-104 cell line?

We bought this cell line 6 months ago and we are trying to make it grow, no success so far. We are using ATCC suggested media and replacing IL-2 every 3 days. Fully media changes (by centrifuging...

13 January 2020 7,719 1 View

EO771 mouse tumor transfection protocol?

Hello! I've been trying to transfect EO771 using lipofectamine 2000, but despite changing the transfection media after 6 hours of incubation I have a high rate of cell death. My untransfected...

13 November 2019 7,883 3 View

How can I calculate Ethanol's boiling point for a specific volume?

08 October 2019 7,724 3 View

How can avoid aggregation of cells in E. Coli staining with ThS and propidium iodide when trying to visualize them in an epifluorescence microscope?

I got competent cells of E. Coli induced with IPTG to express B-amyloid peptide and I stained them with ThS and propidium iodide after permeabilizated with Triton x 100; however, when I want to...

07 August 2019 3,158 1 View

No title

I have used FITC-dextran in my mice via gavage, and collected the blood samples to run together. Should I keep the samples at -20C or 4C? If I freeze my FITC can that affect the resulting...

31 July 2019 137 2 View

How quantify relic DNA in water samples?

I need to quantify the amount of relic/free DNA in water samples from a fish pond. I'm looking for a protocol with these method. Does anyone know of any accurate methodology for this...

21 May 2019 5,636 3 View

Which are the best resources to learn AnyLogic?

17 April 2019 7,086 6 View

Buffer Exchange using column packed with sephadex G-25 effect on protein concentration?

Hello, If i am doing a buffer exchange for an antibody of 1mg/mL, does the elution lose protein in the process of buffer exchange? For example, if i flow through 500 uL of 1mg/mL sample, and...

03 March 2021 6,299 3 View

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 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

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 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

What is the best concentration of cDNA to be used for qPCR?

I have synthesised cDNA from 500ng/ul of RNA. The concentration of the cDNA Is about 5000ng/ul. Should I dilute my cDNA or just use it as it is for my qPCR? I usually use 2ul of cDNA for a 20ul...

24 February 2021 8,618 3 View

Recommendations on homogenizers for RNA isolation?

I will be performing RNA isolation with frozen lung biopsy samples and am in the market for a tissue homogenizer. I am wanting a homogenizer that will fit in a 1.5ml tube. I was looking at the...

24 February 2021 3,623 3 View

RNA-seq DEGs downstream analysis and reporting in literature?

I used EdgeR to get the list of DEGs (differentially expressed genes) from my liver RNA-seq experiment. I have done the gene overrepresentation and GSEA based pathway analysis. I am currently...

23 February 2021 8,969 1 View

Is it efficient to do liquid biopsy to investigate the gene expressions of the tumor?

To do liquid biopsy and look for specific gene expressions there are so many published papers. Thera are different protocols and procedures used to isolate circulating cell free RNA or mRNA from...

23 February 2021 7,326 5 View

RNA degraded after good nanodrop readings?

23 February 2021 8,850 2 View