What kit or method is suitable for the extraction of RNA from around 50 cells?

Dear All, I am wondering if you could recommend a good kit or method for the extraction of RNA from as low as 50 cells. I need to run RNA seq afterwords. Looking forward to your...

31 December 2017 1,411 2 View

Could a tumour suppressor gene enhance the activation of PI3K pathways?

I work on a tumour suppressor gene in breast cancer. The down regulation of this gene in MCF10A cells (using shRNA) caused a 12 folds decrease in the mRNA expression of PI3K catalytic subunit...

31 December 2013 896 8 View

Do I have to confirm the qPCR array data using specific qPCR assays?

I ran qPCR arrays to test the expression of 48 genes in two samples. 10 genes showed significant difference between samples. The assay was very reproducible (nearly same results between...

09 October 2013 3,713 8 View

Is there any way to ship cell lysates (for western and Co-IP) abroad?

The next step of my project is to do western blotting and Co-IP. However, the cells which I will use are not in my lab, they are in my collaborator lab abroad. Because I do not have cell culture...

11 December 2012 4,739 2 View

What are the best ways for selecting genes to be included in qPCR arrays?

I am currently designing qPCR array to investigate the effect of my gene of interest (CSMD1) on the expression of a number of genes. My cell culture studies showed that CSMD1 affects cell shape,...

04 May 2012 1,946 7 View

Does anyone tried mammosphere assay for circulating tumour cells?

01 January 1970 5,309 3 View

What is your perception on using visual analysis or quantitative approaches to evaluate data from single-case research designs (n of 1 trials)?

Results of single-case research designs (i.e., n-of-1 trials) are often evaluated by visually inspecting the time-series graph and computing quantitative indices. A question our research team is...

03 March 2021 687 1 View

How do I increase the yield of my PCR product?

Hello, We would like to increase the yield of our PCR product. We are running a series of PCR reactions that is targeting ~1.1kb sequence. We begin each reaction with ~400pg of template DNA...

02 March 2021 4,029 3 View

Does anyone know HRM channel in a 2 plex QIAGEN Rotor- Gene Q qPCR can be used as a separate channel?

I am going to have 3 different probes in my qPCR work that I am going to do. But I realized that the machine we have in the lab is a Rotor-Gene Q 2plex HRM Platform, saying it has green, yellow,...

01 March 2021 8,544 1 View

How to address this issue in the analysis of Real-Time PCR results?

To dear Researchers, I was analyzing a series of concentration for estimation of Real-Time PCR efficiency. The concentration was 1:10. I used MS-excel to evaluate Slope. The result of slope was -8...

01 March 2021 8,683 4 View

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Does anyone have the experience of using Taq Man probes in the QIAGEN Rotar- Gene qPCR machine?

01 March 2021 5,311 1 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

Genome-wide gene identification protocol ?

Is there any book chapter/book, webpage or research article available to understand genome-wide gene identification?

28 February 2021 8,095 1 View

MiRNA qpcr problem?

Dear All, mirna primer showing some problem in the melting curve? any idea why? As attached is the melting curve. The forward sequence is obtained from miRBase and reverse primer is universal.

28 February 2021 5,008 4 View

What is Flat Lens Antenna?

28 February 2021 5,433 3 View

Any specific pipeline or parameter to analyse total transcriptomics study of brain tissue samples?

I have two groups of brain samples, control and treated for example. It was total RNA nova seq sequencing. I tried all the available pipeline like: star+rsem+deseq2, Hista+stringtie+cuffdiff,...

27 February 2021 356 6 View