Using bbduk for decontaminating sequences? I did Mi-seq for fosmid DNA but got some E. coli sequences contaminating the data, how can I remove them? Someone recommended I use bbduk for this, but I don't know how to do that?Thank you!
I need to separate acetyl co-A, malonyl co-A & free co-enzyme A on TLC, I tried several mobile phases but they didn't work with acetyl co-A, it didn't move. I tried: ethyl acetate, methanol...
06 July 2017 3,599 1 View
Hi, I have a fosmid of size ~40 kb, what would be the best method to sequence it & capture all the insert?
09 October 2016 8,222 3 View
I need to send Fosmid DNA for sequencing, but they told me the concentrations were too low & there are multiple bands. I tried mini-prep Qiagen kit many times & also midi-prep. How can...
07 August 2016 8,161 2 View
(Felczykowska et al., 2014): Overnight cultures from E. coli were centrifuged at 3,500 rpm for 10 min. Afterwards, the cell pellets were re-suspended in 0.01 M Tris-HCl pH 7. Finally the extracts...
05 June 2016 2,529 0 View
What if I don't read the plate immediately, what should I do and how do I preserve the plate? in the fridge or on the bench? and is it reliable to do so?
11 December 2013 5,674 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
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
I am planning to collect human fecal samples for metatranscriptomic analysis using MGI. These samples are from indigenous people living in a region with high temperatures. I will have access to a...
06 August 2024 1,367 3 View