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| Status |
Public on Apr 09, 2024 |
| Title |
Anchor-Enhanced Bead Design for Reduced Oligonucleotide Synthesis Errors in Single-cell sequencing |
| Organisms |
Homo sapiens; Mus musculus |
| Experiment type |
Expression profiling by high throughput sequencing
|
| Summary |
Single-cell transcriptomics, reliant on the incorporation of barcodes and unique molecular identifiers (UMIs) into captured polyA+ mRNA, faces a significant challenge due to synthesis errors in oligonucleotide capture sequences. These inaccuracies, which are especially problematic in long-read sequencing, impair the precise identification of sequences and result in inaccuracies in UMI deduplication. To mitigate this issue, we have modified the oligonucleotide capture design, which integrates an interposed anchor between the barcode and UMI, and a 'V' base anchor adjacent to the polyA capture region. This configuration is devised to ensure compatibility with both short and long-read sequencing technologies, facilitating improved UMI recovery and enhanced feature detection, thereby improving the efficacy of droplet-based sequencing methods.
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| Overall design |
In order to evaluate the accuracy of our spacer UMI assignment we captured mixed mouse and human cells (at ratio of 50:50) and then performed single-cell RNA sequencing.
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| Contributor(s) |
Cribbs AP |
| Citation(s) |
39819888 |
| |
| Submission date |
Apr 08, 2024 |
| Last update date |
Feb 14, 2025 |
| Contact name |
Adam Cribbs |
| E-mail(s) |
adam.cribbs@ndorms.ox.ac.uk
|
| Organization name |
University of Oxford
|
| Department |
NDORMS
|
| Street address |
Windmill Road
|
| City |
Oxford |
| ZIP/Postal code |
OX37LD |
| Country |
United Kingdom |
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| Platforms (2) |
| GPL29439 |
MinION (Homo sapiens; Mus musculus) |
| GPL32819 |
PromethION (Homo sapiens; Mus musculus) |
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| Samples (7)
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| Relations |
| BioProject |
PRJNA1097581 |
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