I want to know exact differences in remodeling events in case of normal translation termination and aberrant translation termination PTC bearing mRNA.
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
I aim to be as skeptical as possible regarding whether a pair of orthologous genes results in the same phenotype in their different but related bacterial organisms under similar environmental...
05 August 2024 6,787 4 View
I am performing ligation of the plasmid and a target gene. The steps I have taken are: 1. Double digestion of the plasmid and target gene 2. Ligation of the plasmid with the target gene 3....
05 August 2024 2,570 3 View
I have an RNA-seq data that I have analysed using Limma-voom and have extracted the gene IDs, log2FC and the p-values. At p value < 0.05, I have over 10,000 DEGs, however, when I run the GO...
31 July 2024 225 2 View
I am currently working on a project involving liposomes and need to determine the maximum volume of siRNA that can be added to a 2.5 mL liposome solution with a total lipid concentration of 10...
30 July 2024 6,420 1 View
I cannot confirm cell surface protein expression by flow cytometry even after transfection and antibiotic selection of cells. Does it take long for proteins to express on the cell surface? the...
28 July 2024 3,178 3 View
I created two potential gene expression cassettes (constitutive and inducible) for expression of a mutant PETase gene on PeptiCloud using the version tree feature, which allows users to create...
28 July 2024 7,559 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
Gene sequencing related trouble shooting
25 July 2024 4,149 2 View
I found that several expression constructs of published protein structures containing an N-teiminal fusion tag GAMGSGIQRPTST. What's the founction of this fusion tag?
23 July 2024 1,353 4 View
Hi Everyone, I'm using an siRNA kit to knock down a target gene. The kit guarantees that the negative control doesn't target any sequence in mouse genome, and when I use BLAST I don't find any...
23 July 2024 2,673 6 View