CUTseq is a versatile method for preparing multiplexed DNA sequencing libraries from low-input samples

Xiaolu Zhang, Silvano Garnerone, Michele Simonetti, Luuk Harbers, Marcin Nicoś, Reza Mirzazadeh, Tiziana Venesio, Anna Sapino, Johan Hartman, Caterina Marchiò, Magda Bienko () and Nicola Crosetto ()
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Xiaolu Zhang: Karolinska Institutet
Silvano Garnerone: Karolinska Institutet
Michele Simonetti: Karolinska Institutet
Luuk Harbers: Karolinska Institutet
Marcin Nicoś: Karolinska Institutet
Reza Mirzazadeh: Karolinska Institutet
Tiziana Venesio: Candiolo Cancer Institute, FPO-IRCCS
Anna Sapino: Candiolo Cancer Institute, FPO-IRCCS
Johan Hartman: Karolinska Institutet
Caterina Marchiò: Candiolo Cancer Institute, FPO-IRCCS
Magda Bienko: Karolinska Institutet
Nicola Crosetto: Karolinska Institutet

Nature Communications, 2019, vol. 10, issue 1, 1-12

Abstract: Abstract Current multiplexing strategies for massively parallel sequencing of genomic DNA mainly rely on library indexing in the final steps of library preparation. This procedure is costly and time-consuming, because a library must be generated separately for each sample. Furthermore, library preparation is challenging in the case of fixed samples, such as DNA extracted from formalin-fixed paraffin-embedded (FFPE) tissues. Here we describe CUTseq, a method that uses restriction enzymes and in vitro transcription to barcode and amplify genomic DNA prior to library construction. We thoroughly assess the sensitivity and reproducibility of CUTseq in both cell lines and FFPE samples, and demonstrate an application of CUTseq for multi-region DNA copy number profiling within single FFPE tumor sections, to assess intratumor genetic heterogeneity at high spatial resolution. In conclusion, CUTseq is a versatile and cost-effective method for library preparation for reduced representation genome sequencing, which can find numerous applications in research and diagnostics.

Date: 2019
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DOI: 10.1038/s41467-019-12570-2

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