How can I analysis ChIP-qPCR data?

I am a beginner of Transcription factor ChIP experiment. I have taken 30ug of protein for each immunoprecipitation and the input was 1%. After immunoprecipitation, my input DNA concentration is...

01 February 2020 1,796 2 View

Which statistical test is best for additive model of genotypic study?

I am planning to do an analysis of both allelic and genotypic level of association of an particular SNP with a disease via case control study.But, I can not understand which statistical test...

07 August 2019 8,182 3 View

What would be the consequences if I take equal ratio of renilla plasmid rather than equal amount in my promoter analysis through luciferase assay?

For my promoter activity analysis, I take 0.12pmol of each different size of construct and 1:100 ratio of renilla vector. I balance the amount of total vector using a filler plasmid. 1) Construct...

02 March 2019 6,455 2 View

Why does site-directed mutagenesis not show specific band in gel?

I am doing site-directed mutagenesis using Inverse-PCR method to mutate a 10bp region of my insert. My vector plus insert size is 5.6kb. I am using Pfu polymerase from NEB to amplify my plasmid...

06 July 2018 2,164 5 View

What should be the minimum difference between your background and vector induced luciferase values?

I am doing Dual luciferase assay in a hard to transfect cell line and got these values- 1) Non-transfected- Firefly 93.16(RLU) and Renilla 123.85(RLU), so F/R is 0.75 2) Empty vector...

04 May 2018 1,981 2 View

What should be the concentration range of my drug to check its activity in cell lines?

I want to check the activity and the efficacy of 5 different drugs in cell lines to evaluate the best one based on their potency and reduced toxic effects. In many papers, researches did take the...

10 November 2017 8,445 11 View

What is Biological replicate in cell culture experiments?

I am a beginner of cell culture experiments and just have started some experiments with Hela cell line.In research papers, it is always highlighted that, each experiment has been done in...

07 August 2017 6,930 2 View

What is true Biological replicate in cell culture experiments?

In my cell culture experiments, first I have revived a vial of cells, subcultured it, then extracted RNA from a particular no of cells for my experiments.After RNA isolation, I have freezed my...

06 July 2017 8,334 1 View

Why did endogeneous controls not show any band in western blot?

I am a beginner of western blot and my target protein if 124kDA whereas the endogeneous controls are 41kDA and 55kDA. I did wet transfer at 100V, 80mins and successfully got the band for target...

05 June 2017 4,613 5 View

Why do the low molecular weight proteins not show any band in western blot?

I am a beginner of western blot and my target protein if 124kDA whereas the endogeneous controls are 41kDA and 55kDA. I did wet transfer at 100V, 80mins and successfully got the band for target...

04 May 2017 9,128 10 View

Can linear DNA be used as template for site-directed mutagenesis?

Is it possible to induce site-directed substitution mutation by quick-change method on linear dsDNA? or it has to be cloned in some vector? If yes, should it be treated with the Dpn1 enzyme...

03 March 2021 401 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

PUC19 as a transfer plasmid for lentiviral delivery?

Hello, Is it possible to use pUC19 as a transfer vector to be packed in using the second generation viral particles packaging system( pMD2.G; psPAX2 plasmids)? As far as I understand it there is...

28 February 2021 4,868 2 View

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28 February 2021 9,936 2 View