Abhay Kanodia

10 Questions 5 Answers 0 Followers

Questions related from Abhay Kanodia

Why my enzyme doesnt have any activity?

Hello all, I am over-expressing acid sphingomyelinase enzyme in HeLa cells. Theenzyme shows expression on western blot but I do not have any activity in in vitro assays. I have checked the...

07 July 2023 396 3 View

How to use miRDeep2?

Hello, I am using the following tutorial for microRNA profiling: https://drmirdeep.github.io/mirdeep2_tutorial.html I was able to follow it until the patching step which is: bash...

01 November 2022 6,818 0 View

Is there a way to compare RNA-seq and scRNA-seq?

I performed RNA-seq and scRNA-seq on the same set of samples but the log2 fold change values are in very different range and I am not sure we can normalize it in any way. Please let me know if...

11 July 2022 6,177 3 View

How to grow MCF10A single cell clones?

Does anyone have experience growing single cell clones of MCF-10A.I have generated gene knockouts and need to pick single cell clones from these cell lines. It is challenging as these cells are...

29 April 2021 1,882 2 View

Does anyone have experience with Blasticidin selection after lentiviral transduction?

I am using MCF10A cell lines. I see different sensitivity to Blasticidin of my control cells (not infected with lentivirus) vs cells infected with lentivirus. I thought this is a good thing as My...

26 February 2021 7,086 1 View

Does antibiotic resistance gene integrate in the genome along with the sgRNA?

In CRISPR mediated cell transduction, does your selectable marker (blasticidin resistance gene) or flourescent protein (gfp or rfp) integrate in the genome? Further, which of the selection...

18 December 2020 9,198 1 View

Why MCF10A cells die after selection with 5ug Blasticidin?

I am making CRISPR knockouts in MCF10A cells and Blasticidin is used as a selection marker. When I add Blasticidin, some cells die which is expected as they haven't received my sgRNA. However,...

12 December 2020 4,735 2 View

Why do I get a lot of colonies upon transformation of digested lenti guide puro?

I digested my lenti guide puro plasmid. The only modification in this plasmid is that it has blasticidin resistance gene instead of puromycin. I get the desired bands at ~8kb but I do not see the...

04 August 2020 3,022 4 View

How do you measure the activity of murine CD8+ T cells after adding anti CD3/anti CD28 beads in vitro?

I have added anti- CD28/anti-CD3 beads to in vitro culture of murine T cells. Some of the cells have increased in size which indicates their activation. However, is there a method to quantify the...

09 August 2019 3,566 7 View

QPCR- (a) Why do I have different melt curves for same primer (b) gene of interest value lower than GAPDH?

Hi, I have attached images for the two questions I have. I use SYBR green for quantification. (a) I have my gene NF2 and I get a different melt curve for different samples. I get a single peak...

01 January 1970 9,310 2 View