What if referance genes are also enfluenced by specific treatment in real time PCR? HOw to analyze data in such situation?

I have found that a specific treatment influences the Ct value of both the reference gene (Actin and Ubiquitin) in RT qPCR experiment. Is it possible to analyze such data?

01 February 2018 9,942 4 View

What is the best concetration of SDS PAGE gel to visualize 10 kDa protein?

Hello, I am facing problem in visualizing protein of 10 KDa size on 15 % SDS PAGE gel. What are the suggestions for solving this problem? Thanks, Suyash

01 February 2018 3,767 3 View

Hello, Anybody working on Pcold GST vector system?

I am unable to induce protein of my interest using pcold GST vector system. I have transformed the vector in BL21 cells. I am unable to see any protein expression on induced culture compared to...

10 November 2017 8,194 6 View

How to calculate annealing temperature for the pcr set up with primers designed for in-fusion cloning?

The primers designed for the infusion cloning system have 15 bp overhang from the vector sequence and around 20 to 25 bp for the sequence from the gene of interest. The question is how to...

04 May 2017 8,987 5 View

Any body have experience using NEB Q5 Site Directed Mutagenesis Kit ?

I am trying to insert 90 bp insertion in 12 KB vector. The insert is a 30 bp tag repeated for 3 times. First I used a 114 bp forward primer and 38 bp reverse primer because there is great chance...

09 October 2016 3,422 11 View

How can I amplify an AT rich genome sequence?

I am trying to amplify an AT rich sequence which is 1543 bp long but still unsuccessful. I have used Phanta Max and Taq enzymes. Please suggest possible solution for it.

06 July 2016 8,966 3 View

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02 March 2021 3,060 3 View

New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before?

Question to you and THEM, the New Journal, "Integrative Psychological and Behavioral Science" -- do you not know, and have you not seen, this done before? There appears to be a core problem for...

02 March 2021 3,024 2 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

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

Could you please recommend some good journal that accept very long revie papers (approximately 6000+ words) in neurosurgery, cancer biology ?

01 March 2021 8,087 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

Where to find data on trees in tropical agroforestry?

Hi, I'm looking for data (mainly related to management: growth rate, canopy size, soil and climate preferences, etc.) about tropical trees used in tropical agroforestry. Have you ever heard about...

28 February 2021 7,356 8 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