Tweet
I was just wondering if anyone has used the minIon for plasmid sequencing? I suppose it might be overkill and a waste or reagents for such a small task. But I am asking because I have large plasmids to sequence (in the 18 to 20 kb range), and the plasmids contain several copies (up to as many as 4 copies) of certain sequences. So using sanger sequencing is costly and labor intensive and we end up with reads that could align to more than one spot on the plasmid making it hard to tell if and where a mutation has occurred. I am also exploring Illumina based methods, but am curious about the nanopore technologies. Seems like a 20 kb plasmid could simply be linearized and the entire molecule sequenced in a single read.
-
-
Have a look at methods for Zika virus sequencing project.
This sounds very similar to zika sequencing project, and should work reasonably well (provided it is medium GC% sequence).
Ideally one would like to make partial digest of the plasmid, aiming to cut most of them once or twice, and make 2D library from it.
PS: keep in mind the ~0.1 - 1% systematic error rate remaining after assembly consensus steps. It would really depend on the actual template sequence.
Hopefully there are no high GC hairpins in the sequence, because they would stop the complementary read (which goes from single stranded template).
So you may have some bits that have very low quality hear the hairpins. -
Hey, did you ever get this working? I am thinking about creating a single cut in the plasmid and sequencing entire AMR plasmid. Know any papers where this has been done?Comment
-
Proteomics platforms are evolving rapidly, with advances in mass spectrometry and affinity-based approaches expanding what researchers can detect and at what scale. As the field moves toward deeper proteome coverage and clinical applications, scientists face an increasingly complex landscape of tools. This article will explore how researchers are navigating these choices to find the right platform for their work.
The systematic characterization of the human proteome has...07-20-2026, 11:48 AM -
Genomics studies in neuroscience face a special challenge due to the brain’s complexity and scarcity of samples. Mapping changes in cell type and state using conventional next-generation sequencing methods remains challenging. Advances in technologies like single-cell sequencing, spatial transcriptomics, and long-read sequencing have opened the door to deeper studies of the brain and diseases like Alzheimer’s, amyotrophic lateral sclerosis (ALS), and schizophrenia.
...07-09-2026, 11:10 AM -
Cancer survival rates have significantly increased in the last few decades in the United States, reaching a combined 70% 5-year survival rate by 2021. Behind this number, there are years of research to find new therapies, drug targets, and early detection methods. But there is one core challenge that keeps slowing down these advances, and it’s about drug resistance.
There is no single reason why many patients don’t respond to treatment as expected. Cancer is...07-08-2026, 05:17 AM
Comment