Cell View

Cell Information


Overall DesignSequencing libraries from cDNA was generated using in-house produced or commercial Tn5 and in a variety of tagmentation buffers and conditions.
SummaryMassively 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.
Dataset viewGSE58652

Samples in HEK293T

Displaying 1-10 of 42 results.
SeriesSampleInstrumentOrganismTitleCell Source
GSE58652GSM1416388Illumina HiSeq 2000Homo sapiensinhouse_0.1pg_rep1HEK293T
GSE58652GSM1416389Illumina HiSeq 2000Homo sapiensinhouse_0.1pg_rep2HEK293T
GSE58652GSM1416390Illumina HiSeq 2000Homo sapiensinhouse_0.1pg_rep3HEK293T
GSE58652GSM1416391Illumina HiSeq 2000Homo sapiensinhouse_100pg_rep1HEK293T
GSE58652GSM1416392Illumina HiSeq 2000Homo sapiensinhouse_100pg_rep2HEK293T
GSE58652GSM1416393Illumina HiSeq 2000Homo sapiensinhouse_100pg_rep3HEK293T
GSE58652GSM1416394Illumina HiSeq 2000Homo sapiensinhouse_10pg_rep1HEK293T
GSE58652GSM1416395Illumina HiSeq 2000Homo sapiensinhouse_10pg_rep2HEK293T
GSE58652GSM1416396Illumina HiSeq 2000Homo sapiensinhouse_10pg_rep3HEK293T
GSE58652GSM1416397Illumina HiSeq 2000Homo sapiensinhouse_1pg_rep1HEK293T

Gene rank in HEK293T