|
Status |
Public on Jun 15, 2020 |
Title |
DNA sequencing for multiple rounds of the pMHC-yeast display selection for 2W, Yae and Fat TCR |
Organism |
synthetic construct |
Experiment type |
Other
|
Summary |
A peptide library was displayed on Ab MHC class II allele expressed on yeast. TCRs of interest were used to select and enrich for cognate peptide-MHC.
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|
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Overall design |
A naive pMHC-yeast display library was selected with biotinylated TCRs of interest present on magnetic beads (Miltenyi) or fluorescently labelled streptavidin. After 4 rounds of TCR selection, DNA was extracted using zymolase for all rounds of selected yeast, enriched for cognate pMHC, as well as the naive library. Isolated DNA/plasmid was amplified by PCR with appropriate barcodes and sequencing was performed using a Miseq.
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|
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Contributor(s) |
Garcia KC, Fernandes RA |
Citation(s) |
32773038 |
|
Submission date |
May 22, 2020 |
Last update date |
Aug 24, 2020 |
Contact name |
Ricardo A Fernandes |
E-mail(s) |
ralexandrelf@gmail.com
|
Organization name |
Stanford University
|
Department |
Molecular and Cellular Physiology
|
Lab |
KC Garcia Beckman Bldg
|
Street address |
279 Campus Drive
|
City |
Stanford |
State/province |
California |
ZIP/Postal code |
94301 |
Country |
USA |
|
|
Platforms (1) |
GPL17769 |
Illumina MiSeq (synthetic construct) |
|
Samples (2) |
|
Relations |
BioProject |
PRJNA634545 |
SRA |
SRP262990 |
Supplementary file |
Size |
Download |
File type/resource |
GSE151070_2w_20161109_uniques_stats.csv.gz |
9.3 Kb |
(ftp)(http) |
CSV |
GSE151070_2w_Round3.pfam.txt.gz |
5.3 Mb |
(ftp)(http) |
TXT |
GSE151070_2w_Round4.pfam.txt.gz |
7.2 Mb |
(ftp)(http) |
TXT |
GSE151070_Fat_Round3.pfam.txt.gz |
5.9 Mb |
(ftp)(http) |
TXT |
GSE151070_Fat_Round4.pfam.txt.gz |
7.2 Mb |
(ftp)(http) |
TXT |
GSE151070_Fat_sclean_uniques_stats.csv.gz |
30.9 Kb |
(ftp)(http) |
CSV |
GSE151070_effector1_Round3.pfam.txt.gz |
6.0 Mb |
(ftp)(http) |
TXT |
GSE151070_effector1_round4.pfam.txt.gz |
1.4 Mb |
(ftp)(http) |
TXT |
GSE151070_mediummay_naive.fasta.gz |
22.5 Mb |
(ftp)(http) |
FASTA |
GSE151070_readme.xls |
32.0 Kb |
(ftp)(http) |
XLS |
SRA Run Selector |
Raw data are available in SRA |
Processed data are available on Series record |