Back to Search
Start Over
CUTseq is a versatile method for preparing multiplexed DNA sequencing libraries from low-input samples.
- Source :
-
Nature communications [Nat Commun] 2019 Oct 18; Vol. 10 (1), pp. 4732. Date of Electronic Publication: 2019 Oct 18. - Publication Year :
- 2019
-
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.
- Subjects :
- A549 Cells
Cell Line, Tumor
DNA isolation & purification
DNA metabolism
DNA Restriction Enzymes metabolism
HeLa Cells
Humans
MCF-7 Cells
Reproducibility of Results
DNA genetics
Gene Library
High-Throughput Nucleotide Sequencing methods
Paraffin Embedding methods
Sequence Analysis, DNA methods
Subjects
Details
- Language :
- English
- ISSN :
- 2041-1723
- Volume :
- 10
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature communications
- Publication Type :
- Academic Journal
- Accession number :
- 31628304
- Full Text :
- https://doi.org/10.1038/s41467-019-12570-2