Somatic mutations - how to validate NGS data?

Hi, I'm searching for methods to validate somatic mutations in pancreatic beta cells. The mutations are found by Next Generation Sequencing and I have try Sanger Sequencing with no results.

12 March 2018 1,821 5 View

Glucose stimulation of rat beta cells for insulin measurement?

Can anyone suggest a protocol for glucose stimulation of rat beta cells for insulin measurement. I can see final glucose concentration as 16,7mM but I dont know why?

26 February 2018 4,742 3 View

Best way to inactivate RNase III-A?

I require 100 uL of bovine ribonuclease III-A stock solution (5mg/mL). The product from Sigma-Aldrich is a very fluffy substance. I need to inactivate this twice at 100 degree C twice. Does anyone...

31 January 2017 1,632 3 View

Normalization of small RNA-seq with housekeeping RNA?

I am working on some small RNA-Seq data and trying to normalize them. From literature, I found that people are using some housekeeping small RNA as reference for the normalization in qPCR. Can...

01 November 2016 4,047 3 View

What is production mechanism of Haft Kel field?

how is Haft Kel Field produced and how is the CO2 as injection considered in the flow simulation?

09 July 2016 882 2 View

Shelf life of TRIS?

I just started in a new lab and am about to start running western blot. However, the TRIS powder they have here is from 2002 and I'm a bit reluctant using it. Does anyone have experience using old...

26 April 2016 6,451 5 View

Evolution of ageing?

Recently some papers have shown ageing cells to be part of development. If ageing has a function in development, then ageing may be adaptive. Or?

19 January 2014 5,355 17 View

Two questions: using fibronectin in transwell migration assays?

I've been having a lot of trouble trying to get my assays to work. I'm concerned that all I'm seeing is background migration. It's just B16BL6 melanoma cells (usually 1 x 10^5), using 10% FBS as a...

14 May 2013 5,684 15 View

Is possible to publish a phage draft genome as announcement in a journal (e.g. J virol or J Bact) or must you have the complete sequence?

What is the minimal genomic information to required to be accepted for publishing?

21 March 2012 4,643 1 View

What is the most Operating System (OS) which can be INSTALL on Tiny hard devices like Raspberry Pi (RPi) hardware ?

I'm targeting to deploy a mesh network and manually configure MANET routing protocols. I'm preparing scenarios, architectures, and hard devices needed to do that. Are there some step-by-step...

03 March 2021 1,931 5 View

Which of the Covid-19 Vaccines would elicit the best immunoprotection and why?

I want to know which of the available Covid-19 Vaccines would elicit the best immunological response and why

02 March 2021 1,388 3 View

No title

02 March 2021 3,060 3 View

How can I integrate a routing protocol in a tiny device (e.g. Raspberry Pi)?

Hi Community, I'm facing the issue of integration/compiling a new routing protocol in a WSN simulator. The final goal is to successfully add, configure this routing protocol in hardware devices...

01 March 2021 9,332 6 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