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.
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 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 want know exact mechanistic model of EJC, Upf proteins triggering NMD.
09 October 2013 6,739 2 View
Hello, I am currently having problems with RNA extraction. I am using mouse liver (C57BL6J), and I have extracted RNA from mouse liver before. Before this experiment, my final RNA pellets were...
11 August 2024 7,082 3 View
I have reverse sequences (AB1 format), can I base on reverse DNA sequences to perform nucleotide alignment, convert nucleotides to amino acids and deposit the sequence in GenBank database?
11 August 2024 5,138 1 View
I have been doing the m6A dot blot for a while with no improvement, I am extracting the RNA, and I can see the dots although the three biological replicas give a different reading on the memberan...
10 August 2024 8,539 5 View
I'm cloning a fragment of 3200 nts into plasmid. The cloning was successful, however, 02 amino acids were mutated. Now I want to fix these 02 aa by site-directed mutagenesis technique using...
08 August 2024 4,645 2 View
I've been performing RNA extraction on cotton petiole tissue for a few months now using the method described in the following paper, a derivative of the typical hot borate method...
08 August 2024 9,882 2 View
After performing symmetric PCR, PCR purification was performed. Afterwards, asymmetric PCR was performed using the PCR purification product as a template, but no ssDNA band was confirmed in the...
08 August 2024 1,668 3 View
I'm trying to find a DNA extraction method for fungi that does not require equipment and heating. Is there anyone who can suggest an alternative option? Thank you
08 August 2024 4,733 2 View
I am currently working on LncRNA; to know the lncRNA-protein interactions I want to do RNA pull down assay, so I need to design primers with T7 promoter. I need assistance in this regard.
07 August 2024 6,622 1 View
Hi! So i attempted to understand a novel protein behavior towards heat application by analyzing its secondary structure change. I subjected the protein to a thermal denaturation analysis using...
06 August 2024 1,989 3 View
Recently, we observed that 99% of the sequences in our RNA-seq data corresponded to the E. coli genome. Despite multiple DNAse treatments after RNA extraction and ribosomal depletion, we were...
06 August 2024 807 3 View