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You can also try a 95C denaturation (instead of the usual NaOH denaturation). 1 minute at that temperature (in RSB or EB should be fine), just make sure you have a decent volume there so you don't evaporate everything. TruSeq Custom Amplicon product does this method to make the libraries nice and single stranded for cluster generation. -
Anecdotally we find MiSeq to be the best sequencer for odd samples. You best bet is to sequence this library there. You can always a run a nano flowcell at a much reduced cost to test before you run full library.Leave a comment:
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Library with strong secondary structures
What is known about the strand displacement activity of the DNA polymerase Illumina uses for bridge amplification? I have a library with strong secondary structures and I am worried that clusters do not form well due to the secondary structures. Does anyone have any tips to improve cluster formation for such a library?
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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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