This is indeed concerning. We synthesized our in-line barcoding reagents to have the first 4 bases be randomized followed by our less complex sequences.
Let us all know if you have any issues going forward with the 4 base randomization and the new RTA?
-Tom Blomquist
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Hi Nickp,
A little bit late but 2.8 sounds wrong, the current versions are only 1.9
cheers.Leave a comment:
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RTA v2.8 : Conflicts with low complexity sequence
We routinely use an in-line barcoding system with bases 1-4 as variable barcode nucleotides and base 5 is a mandatory T in all indexed libraries.
We have run several such libraries under RTA 2.6 using a PhiX lane to call thresholds. An Illumina tech support suggested this might now be a problem with RTA v2.8 as it used the first 5 nucleotides to threshold rather than the first 4 that RTA2.6 used. Even with a PhiX lane for thresholding they are worried that our low complexity start to all our reads might now cause a conflict.
Anyone else using low complexity reads in RTA 2.8 ?. Any problems over RTA 2.6 ?
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The first FDA-approved CRISPR-based therapy marked the transition of therapeutic gene editing from a dream to reality1. CRISPR technologies have streamlined gene editing, and CRISPR screens have become an important approach for identifying genes involved in disease processes2. This technique introduces targeted mutations across numerous genes, enabling large-scale identification of gene functions, interactions, and pathways3. Identifying the full range...08-27-2024, 04:44 AM
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