I need a tested RNA dot blot protocol!

I am doing RNA dot blot experiment to probe epitranscriptomic changes in brain tissue. I am not getting good signal with total RNA. I doubt my denaturation step as I am heating my samples at 95...

06 July 2017 9,436 1 View

Why do I get a contamination in my H1 cells?

I am working on H1 human embryonic stem cells and culturing them on matrigel coated plates. On the second day of passage, I could see the media appearing cloudy and plate looks grainy with a lot...

06 July 2015 3,586 8 View

Any suggestion on a problem in sub-culturing H1 human embryonic stem cells with feeder free culture conditions?

I am working on H1 cells, I am sub-culturing in 10 cm dish coated with matrigel at 1:10 split ratio of confluent cells. I am having this frequent problem of cells not getting colonized on 3rd of...

05 June 2015 6,906 2 View

Does anyone have a composition of manually made TRIZOL reagent that worked well?

I am trying to isolate RNA from THP1 cell line using manually made TRIZOL reagent, I am getting good intact RNA but along with it I am getting huge DNA contamination. So I want to somehow minimise...

07 August 2014 5,036 8 View

Which is the most reliable method for quantification of gene expression data in qPCR experiment?

There are many methods for quantification of gene expression (fold change calculations) through real time PCR such as Pfaffl method, delta ct method, standard curve method. So I want to know which...

07 August 2014 2,115 4 View

What is the best reference gene for normalising the expression level of a given gene of interest in qPCR experiment?

07 August 2014 5,032 7 View

How does the interaction between eRF3 and PABP stabilize mRNA during translation termination?

mRNA stabilization as a result of translation termination.

10 November 2013 7,737 2 View

How does the interaction of exon junction complex (EJC) and Upf proteins with translation termination factors lead to mRNA decay?

I want to know the reason why interactions between translation termination factors and EJCs lead to mRNA decay during NMD.

10 November 2013 6,120 0 View

What is the utility of real time PCR over northern blot?

Utility of real time pcr over northern blotting

10 November 2013 6,143 3 View

How Exon Junction Complex (EJC) together with Upf proteins trigger non-sense mediated mRNA decay?

I want know exact mechanistic model of EJC, Upf proteins triggering NMD.

09 October 2013 6,696 2 View

Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence?

I am on the lookout for the Enhanced Yellow Fluorescent Protein (Aequorea victoria) DNA sequence. Does anyone know where I can find it? Thank you in advance

03 March 2021 3,568 1 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

Gel electrophoresis result for total RNA samples, 1.5 Agarose gel was used, How to improve the integrity of RNA?

Gel electrophoresis, RNA degradation, RNA extraction from fresh tissue

02 March 2021 5,433 5 View

How to quantitative analize the data of TNBS assay for gelatin?

Estemeed colleagues, I found some issues regarding the quantification of the data for TNBS assay. There are different protocols on how to perform that but it is clear to me the "fil rouge" that...

02 March 2021 2,616 1 View

Sample dilution for quantitative analysis using ELISA?

If the detection range is in ng/ml but the reference range is in ug/ml for a molecule or protein in serum or plasma .how to dilute and what is the initial volume to be taken for quantitative analysis

02 March 2021 7,670 3 View

Do you know good and user-friendly programs to measure plant quantitative traits?

For my research i will need to measure plant quantitative traits (especially leaves area and roots length, but would be nice to add some more information). I recently discovered...

01 March 2021 5,035 2 View

Does anyone have a recommendation for yeast reporter gene plasmid/protocol?

Hi, could anyone recommend a plasmid and/or protocol for reporter gene assay in S. cerevisiae? I want to assess the effect of growth conditions on a transcription factor, so I want to clone it`s...

01 March 2021 210 1 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

Samples to study circular dichroism (chirality)?

I want to build a low-cost polarimeter to study circular dichroism (chirality). What type of samples can I use? I am looking for samples that can be found outside the laboratory.

01 March 2021 9,101 3 View