Seeking to learn the mRNA circularisation significance.
Check this one
http://www.ncbi.nlm.nih.gov/pubmed/12575997
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 9,488 1 View
I have seen many papers performing divergent PCR to detect the circular RNA products. I am not sure if the reverse transcription of circular RNA yields circular DNA with random hexamers.
05 June 2017 9,487 0 View
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 3,641 8 View
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 6,956 2 View
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 5,106 8 View
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 2,192 4 View
I was trying to analyse expression levels of certain genes after treatment of monocytic THP1 cell line with antigenic proteins through qPCR and i tried using the reference genes beta actin and...
07 August 2014 5,082 7 View
mRNA stabilization as a result of translation termination.
10 November 2013 7,788 2 View
I want to know the reason why interactions between translation termination factors and EJCs lead to mRNA decay during NMD.
10 November 2013 6,166 0 View
Utility of real time pcr over northern blotting
10 November 2013 6,204 3 View
What is the most suitable experimental setting to understand the consequences of a particular gene: (1) targeted degradation of the specific transcription factor (TF) of the gene, followed by...
04 August 2024 6,265 1 View
How much volume of siRNA can I add to 2.5 mL of liposomes with a total lipid concentration of 10 mg/mL?
27 July 2024 6,920 1 View
Hello! We are using LNP to deliver circular mRNA into T cells. However, compared with linear mRNA, the transfection efficiency and protein expression duration are similar. Are LNP generation or...
21 July 2024 4,322 1 View
What happens in snapTotal- seq? (Single-cell Total-RNA Profiling Unveils Regulatory Hubs of Transcription Factors https://www.nature.com/articles/s41467-024-50291-3) In this protocol, several RT...
18 July 2024 10,010 0 View
I am treating human cells with an AAV vector encoding for my gene of interest tagged by FLAG. I want to quantify the mRNA expression of my gene of interest via qPCR to discriminate the endogenous...
14 July 2024 8,482 3 View
Hello, I'm generating stable cell lines by transfecting plasmid DNA with lipofectamine 3000. The cells went through antibiotics selection for 3 weeks (polyclonal selection) and when I analyzed...
11 July 2024 4,812 5 View
Hello, I discovered that a compound that I use, which get integrated into RNA, might have an unspecific impact on mRNA stability of a gene of interest. To confirm that, I pretreated my cells for...
10 June 2024 276 2 View
Hi, I am very new to Bioinformatics. Recently, I have the project that aims to perform taxonomic anlysis on raw reads from mixed samples taken from environment. For example, we perfromed NGS...
09 June 2024 6,537 3 View
Hi all, Last week, I seeded cells from the same sample into 4 wells (fibroblasts; 300000 cells/well; 1ml/well). well-1 and well-2 are for qRT-PCR test; well-3 and well-4 are for Western blot...
09 June 2024 2,482 3 View
I am using mouse bone marrow derived MSCs (passgae 3-6) for my in vitro cytokine priming experiment. My basal media is 2% FBS-DMEM. I am using 10 ng/ml TNF-a, or IFN-g or both for 24 hours. I see...
27 May 2024 8,935 0 View
