What is the meaning of zero or undetectable gene expression ct value of RT-PCR?

I'm getting zero or no ct value in my gene expression analysis during RT-PCR analysis. I'm not understanding the meaning of zero ct value, how I should interpret the data now. Should I consider...

02 March 2020 2,976 3 View

Is it ok to use transparent 96 wells plate for h2dcfda assay or I should use black plate with transparent base?

I'm using transparent polystyrene 96 well plate for H2dcfda assay, is it ok or I should use black plate with transparent base.

01 February 2020 5,212 3 View

Which statistical test to perform my MTT dataperformed for different time intervals with different concentrations of a bacterial toxin?

I have treated THP1 and AGS cells for 12, 24 and 48 hours with a bacterial toxin concentrations 0, 5, 10, 20, 40, and 80 ug/ml. Now I want to prove my results with statistical methods but I'm...

10 November 2019 6,437 6 View

Problem during pET15b and pET22b restriction digestion with XhoI and NdeI?

Hello everyone, I'm digesting 2 ug of midiprep isolated pET15b and 22b with BamHI and XhoI at 37 oC for overnight. There is smearing in gel and size of the cut vectors also not appearing actual,...

05 June 2019 1,520 4 View

What would be the conductivity of Tris buffer having molar concentrations 1.5 and 0.5 M with respect to pH 8.5 and 6.5 respectively?

I'm preparing tris buffers of molarity 1.5 and 0.5 M with respect to pH 8.5 and 6.5 respectively. The conductivity of molar concentration 1.5 is 26.0 ms/cm and for 0.5 M is 33.6 ms/cm. Is it a...

03 April 2019 6,071 9 View

How to write short name for downloaded protein sequences quickly to save time?

I have downloaded 1000 protein sequences having very big name in FASTA format. I want to cut short these name in a quick manner. Is it possible through and command line or software or I will have...

03 April 2019 7,274 4 View

Is it good to strip gaps in a protein MSA file for phylogenetic analysis?

I'm analyzing 445 protein sequences of a single protein for phylogenetic analysis. Multiple sequence alignment of these sequences is showing gaps of different sizes. Is it ok to keep these gaps...

03 April 2019 8,456 3 View

How many colonies we should expect after transformation of ligation mixture during a cloning experiment?

I'm getting more than 200 colonies after transforming 2 ul of my ligation mixture from 1;3 ratio. I used 100 ng of vector and now it is going hectic to screen a positive clone because the colies I...

01 February 2019 8,673 4 View

Reason for long streaky colonies after transformation?

Hello every one, I'm getting long streaky colonies after transforming pET15b in DH5alpha. Kindly suggest the reason.

01 February 2019 6,552 7 View

What would be best competent cell to maintain and expression of a secreted Gram negative bacterial toxin gene?

I'm cloning a secreted Gram negative bacterial toxin and not finding any colonies with DH5alpha and pET15b. I'm just wondering what could be the best competent cell for keeping and expression of...

31 December 2018 8,914 8 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

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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

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

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28 February 2021 9,936 2 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