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This is an interesting thread - we also had a drop in MiSeq performance (in low complexity libraries) after upgrading to 2.6 and like some of you rolled back to 2.5. I was told there was nothing in the upgrade that might cause this but our runs turned to custard about the time we upgraded. I am staying put in 2.5 for as long as possible.
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Hi GenoMax, not in so many words, but they had reports of poor performance and the primary improvements in MCS 2.6 were for the Trusight Tumor 15 workflow, so rolling back was suggested if we weren't running that.Comment
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Just in case you haven't seen it: The 2.6 version is currently not available due to a "review"
From: https://support.illumina.com/downloa...s_updater.htmlThe recently-released MiSeq Updater v2.6.1.4 has been temporarily removed from our website for review. We will update customers as information becomes available.Comment
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Well it's clearly library specific, though I can't figure out how. I had a successful run last week 75% 7pM 16s amplicons, 20% 8pM truseq genome, 5% phiX. Tried running the problem library again-60% 6pM 16s amplicon, 10% ITS amplicon, 15% 8pM nextera genome, 15% phiX. Again the first read and the indices look great (700k, 86%PF, 16% aligned to phiX). Second read is horrible and only 3% aligned to phiX. This run is higher diversity (though still very low diversity by general Illumina guidelines) and lower clustering than the last successful run.Last edited by thermophile; 02-10-2016, 02:19 PM.Microbial ecologist, running a sequencing core. I have lots of strong opinions on how to survey communities, pretty sure some are even correct.Comment
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This run finally worked on a different machine. FAE is coming out next week to check the temps on both machines and to adjust my machine to match the one that worked.Microbial ecologist, running a sequencing core. I have lots of strong opinions on how to survey communities, pretty sure some are even correct.Comment
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@Number6: We finally hit the "lost focus" error late in cycle on one of runs and were advised by tech support to roll back to MCS v.2.5.x. So far so good.Comment
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My temp control module is bad!! It would hit a temp but then not hold it. FAE is here replacing itMicrobial ecologist, running a sequencing core. I have lots of strong opinions on how to survey communities, pretty sure some are even correct.Comment
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Yet another run failed (after the TCM was replaced). Second level support suggested adding more primers since the TM of the primers (v4 bacteria, aka the ones that most microbiome people use) is low 62-66. This seems to have fixed the issue. I'll see how long this fix lastsMicrobial ecologist, running a sequencing core. I have lots of strong opinions on how to survey communities, pretty sure some are even correct.Comment
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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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