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Series GSE58652 Query DataSets for GSE58652
Status Public on Jul 14, 2014
Title Tn5 transposase and tagmentation procedures for massively scaled sequencing projects
Organisms Homo sapiens; Mus musculus; synthetic construct
Experiment type Expression profiling by high throughput sequencing
Summary Massively parallel DNA sequencing of thousands of samples in a single machine-run is now possible, but the preparation of the individual sequencing libraries is expensive and time-consuming. Tagmentation-based library construction, using the Tn5 transposase, is efficient for generating sequencing libraries but currently relies on undisclosed reagents, which severely limits development of novel applications and the execution of large scale projects. Here, we present simple and robust procedures for Tn5 transposase production and optimized reaction conditions for tagmentation-based sequencing library construction. We further show how molecular crowding agents both modulate library lengths and enable efficient tagmentation from sub-picogram amounts of cDNA. Comparison of single-cell RNA-sequencing libraries generated using produced and commercial Tn5 demonstrated equal performances in terms of gene detection and library characteristics. Finally, as naked Tn5 can be annealed to any oligonucleotide of choice, for example molecular barcodes in single-cell assays or methylated oligonucleotides for bisulfite sequencing, custom Tn5 production and tagmentation enables innovation in sequencing-based applications.
Overall design Sequencing libraries from cDNA was generated using in-house produced or commercial Tn5 and in a variety of tagmentation buffers and conditions.
Contributor(s) Picelli S, Björklund Å, Sandberg R, Reinius B, Winberg G
Citation(s) 25079858
Submission date Jun 19, 2014
Last update date May 15, 2019
Contact name Rickard Sandberg
Organization name Karolinska Institutet
Department Department of Cell and Molecular Biology
Street address Berzelius vag 35
City Stockholm
ZIP/Postal code 17177
Country Sweden
Platforms (3)
GPL11154 Illumina HiSeq 2000 (Homo sapiens)
GPL13112 Illumina HiSeq 2000 (Mus musculus)
GPL15228 Illumina HiSeq 2000 (synthetic construct)
Samples (67)
GSM1416385 generuler1
GSM1416386 generuler2
GSM1416387 generuler3
BioProject PRJNA253099
SRA SRP043417

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Supplementary file Size Download File type/resource
GSE58652_RAW.tar 22.5 Mb (http)(custom) TAR (of TXT)
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Raw data are available in SRA
Processed data included within Sample table

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