Forgot to change qPCR reaction volume?

Hi, a quick question? I just shifted to 10ul from our regular 25ul PCR protocol. I used SYBR Green in BioRad CFX. During set up the program, I forgot change the reaction volume from 25 to 10? I...

09 October 2019 2,911 3 View

Has anyone used Hoechst staining for measure DNA on 6 well plates?

I am planning to measure DNA concentration of fixed mammalian cells on 6 well plates. I am using Hoescht for staining. Do I need to use any cell permeabiling agent before I stain them? Should I...

09 October 2018 4,053 3 View

My negative control have comet tails, why?

I'm treating my CHO cells with 100uM H202 for positive control. cells are dissociated by 20mM EDTA in 1X PBS. Trying to do all procedure under low light. 1% Low melting agarose temperature was...

02 March 2018 4,229 14 View

About confusing term: Intron, exon and nonfunctional or non-coding regions of genome.

About confusing term: Intron, exon and nonfunctional or non-coding regions of genome. Are introns and the nonfunctional regions are same?

08 September 2012 5,694 15 View

About Bioinformatics....

Hi, I'm very new in this field. Can anybody tell me where should I start..??? Anybody pls suggest me some paper, from where I can understand what types of work is going on in this field..... I...

10 November 2011 647 17 View

Cassette PCR

Hello everybody, I'm a new member of this group. I've recently come to known about the term 'Cassette PCR'. But I couldn't gather enough information about it. Can anybody help me...???

05 June 2011 8,024 2 View

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02 March 2021 3,060 3 View

What analysis to choose with a large neste dataset and clearly skewed or kurtotic distributions?

Hello! I have a dataset of n=3000 nested within 8 countries with approximately 200 or 400 responses in each country. I originally planned to perform multilevel modelling with 4 dependent variables...

02 March 2021 6,865 1 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

Interested to stain mice tissue with WGA 488 conjugate, anybody has a protocol to recommend?

Interested to stain mice brain tissue with WGA 488 conjugate, appreciate if anyone willing to offer me some guidance or suggestions! Thank you!

28 February 2021 3,951 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

No title

28 February 2021 9,936 2 View