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I am trying to making Illumina sequencing library using transposon based method to add adaptor. However, instead of using the adaptor used in Nexera kits, I would like to use the Y-shaped adaptor. Thus, the gap between the bottom stand of adaptor and bottom strand of genomic DNA need to be repaired. I want to find a method to do gap filling, however, I do not want the DNA to the 3' of the gap (i.e., bottom strand of the adaptor) to be removed by 5'-3' exonuclease activity of DNA Pol I and then re-synthesized. Can I use Klenow fragment here? It is also strange that no one seems to use a Y-shaped adaptor in transposon based library prep. Is there any technical difficulties here?
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The field of epigenetics has traditionally concentrated more on DNA and how changes like methylation and phosphorylation of histones impact gene expression and regulation. However, our increased understanding of RNA modifications and their importance in cellular processes has led to a rise in epitranscriptomics research. “Epitranscriptomics brings together the concepts of epigenetics and gene expression,” explained Adrien Leger, PhD, Principal Research Scientist on Modified Bases...Yesterday, 07:01 AM -
Proteins are often described as the workhorses of the cell, and identifying their sequences is key to understanding their role in biological processes and disease. Currently, the most common technique used to determine protein sequences is mass spectrometry. While still a valuable tool, mass spectrometry faces several limitations and requires a highly experienced scientist familiar with the equipment to operate it. Additionally, other proteomic methods, like affinity assays, are constrained...04-04-2024, 04:25 PM
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