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Status |
Public on Feb 09, 2016 |
Title |
High-throughput measurements of fitness effects of mutations in the yeast U3 snoRNA |
Organisms |
Saccharomyces cerevisiae; synthetic construct |
Experiment type |
Other
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Summary |
We used a molecular barcoding approach to quantify the fitness of ~60,000 randomly mutated variants of the Saccharomyces cerevisiae U3 snoRNA (the data were further used to calculate a map of intragenic epistatic interactions within this RNA molecule, as descibed in Puchta et al. 2016)
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Overall design |
We generated two independent mutant libraries ('Small' with ~3 and 'Big' with ~10 SNPs per allele) on centromeric plasmid, with 20nt random barcodes placed in a non-transcribed region downstream of the gene (PE Samples 30-37 - paired-end sequencing of: U3, 70nt linker and 20nt barcode); next we performed competition experiments in several conditions (E1-5) on populations of yeast cells transformed with both libraries and measured frequencies of mutated variants in time-points during the experiment by deep sequencing of barcodes (Samples 1-29).
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Contributor(s) |
Kudla G, Puchta O |
Citation(s) |
27080103 |
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Submission date |
Feb 08, 2016 |
Last update date |
May 15, 2019 |
Contact name |
Olga Anna Puchta |
E-mail(s) |
olga.puchta@igmm.ed.ac.uk
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Organization name |
University of Edinburgh
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Department |
MRC Human Genetics Unit IGMM
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Lab |
Greg's Kudla Lab
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Street address |
Crewe Road
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City |
Edinburgh |
ZIP/Postal code |
EH4 2XU |
Country |
United Kingdom |
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Platforms (3) |
GPL17143 |
Illumina MiSeq (Saccharomyces cerevisiae) |
GPL19604 |
Illumina HiSeq 2500 (synthetic construct) |
GPL21440 |
Ion Torrent Proton (synthetic construct) |
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Samples (37)
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Relations |
BioProject |
PRJNA311244 |
SRA |
SRP069793 |