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It's possible that the double peak is caused by running too much sample for the chip (I'm guessing your image is from a Bioanalyzer chip or something similar). We've seen some seriously bizarre peaks on our Bioanalyzer (some double like yours and one that was actually a triple peak) that are remedied simply by re-running the sample at a 1:5 or 1:10 dilution.
Sometimes this eliminates all trace of a high MW smear as well, but typically I would guess that the presence of high MW fragments is due to over-amplifying the library. Do you usually amplify for 18 cycles? That is a little high in my experience (which, granted, I don't have tons of yet... ), but if it normally works for you than it might not be the issue.
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by seqadmin
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...-
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03-22-2024, 06:39 AM -
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by seqadmin
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03-08-2024, 10:41 AM -
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