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  HEK293T

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
PMID25079858

Samples in HEK293T

Displaying 21-30 of 42 results.
SeriesSampleInstrumentOrganismTitleCell Source
GSE58652GSM1416408Illumina HiSeq 2000Homo sapiensinhouse_low_1pg_rep2HEK293T
GSE58652GSM1416409Illumina HiSeq 2000Homo sapiensinhouse_low_1pg_rep3HEK293T
GSE58652GSM1416410Illumina HiSeq 2000Homo sapiensinhouse_low_1pg_rep4HEK293T
GSE58652GSM1416411Illumina HiSeq 2000Homo sapiensinhouse_low_500pg_rep1HEK293T
GSE58652GSM1416412Illumina HiSeq 2000Homo sapiensinhouse_low_500pg_rep2HEK293T
GSE58652GSM1416413Illumina HiSeq 2000Homo sapiensinhouse_low_500pg_rep3HEK293T
GSE58652GSM1416414Illumina HiSeq 2000Homo sapiensinhouse_low_500pg_rep4HEK293T
GSE58652GSM1416437Illumina HiSeq 2000Homo sapiensXT_0.1pg_rep1HEK293T
GSE58652GSM1416438Illumina HiSeq 2000Homo sapiensXT_0.1pg_rep2HEK293T
GSE58652GSM1416439Illumina HiSeq 2000Homo sapiensXT_0.1pg_rep3HEK293T

Gene rank in HEK293T

Displaying 21-30 of 19442 results.
Rank orderGene SymbolEnsembl ID
21RPL36ENSG00000130255
22ZNF84ENSG00000198040
23RPS4XENSG00000198034
24PRDX1ENSG00000117450
25HIST1H4CENSG00000197061
26TPI1ENSG00000111669
27PPP1R11ENSG00000204619
28H2AFZENSG00000164032
29RPL5ENSG00000122406
30TUBA1BENSG00000123416