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Has anyone observed that when making paired-end libraries the yield of final PCR product is much less as compared to making single-read libraries? I have made libraries in parallel from the exact same nebulized DNA using both kits (repeated twice, using 2 different paired end kits) with the same result. The concentration of PCR product is more than double in the single read library vs the paired end. I suspect the problem is the PCR step, assuming the efficiency of the polishing/tailing/ligation shouldn't be any different for the two kits. Has anyone tried to optimize the PCR beyond the standard protocol?
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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 -
Despite advancements in sequencing platforms and related sample preparation technologies, certain sample types continue to present significant challenges that can compromise sequencing results. Pedro Echave, Senior Manager of the Global Business Segment at Revvity, explained that the success of a sequencing experiment ultimately depends on the amount and integrity of the nucleic acid template (RNA or DNA) obtained from a sample. “The better the quality of the nucleic acid isolated...03-22-2024, 06:39 AM
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