DNA shearing by french press?

Is it possible to shear DNA by french press to get fragment size about 500 bp?

08 October 2017 1,014 12 View

Prediction tool for DNA-binding site?

Hi, could you recommended me any prediction software for DNA-binding sites of my protein? I have protein with predict structure using Psipred (pdb file) and fasta file with genome. I would like to...

26 July 2017 7,171 3 View

Denaturing PAGE of DNA - how to get sharp bands?

Our research group has recetly bought CBS Scientific Adjustable Sequencing Kit (SG-400-20) and we are facing troubles with our electrophoretic separations because we are unable to obtain sharp...

26 February 2017 4,223 3 View

How to balance ChIP-seq library prep for multiplex sequencing?

I am preparing ChIP-seq libraries for Illumina multiplexing. I have large amount of the ChIP-input DNA compared to the ChIP-output. I wonder if it is better to keep the DNA amount for library...

16 August 2016 1,075 4 View

RNA immunoprecipitation - why Western blot doesn't work but RT-PCR does?

Hi everyone! I try to optimize immunoprecipitation method followed by RNA isolation and RT-PCR/real-time. I did it for a first time and I did not have any experience with this. I managed to...

02 August 2016 2,013 10 View

Has anyone experience with ChIP and bacterial transcriptional factors?

I am going to try this method for prokaryotic cells - gram negative bacteria. I want to define regulon of DNA-binding protein. My protein is non specific binding to dsDNA. Is it a problem?

07 February 2016 7,323 3 View

Can we classify HV and LV lines based on line length?

I have dataset which shows the length of power lines. I need to classify the lines based on the line length. Is there a rule to classify the High voltage (HV) and low voltage (LV) lines based on...

03 March 2021 4,116 4 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 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

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 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