Abstract：
:Despite developments in targeted gene sequencing and whole-genome analysis techniques, the robust detection of all genetic variation, including structural variants, in and around genes of interest and in an allele-specific manner remains a challenge. Here we present targeted locus amplification (TLA), a strategy to selectively amplify and sequence entire genes on the basis of the crosslinking of physically proximal sequences. We show that, unlike other targeted re-sequencing methods, TLA works without detailed prior locus information, as one or a few primer pairs are sufficient for sequencing tens to hundreds of kilobases of surrounding DNA. This enables robust detection of single nucleotide variants, structural variants and gene fusions in clinically relevant genes, including BRCA1 and BRCA2, and enables haplotyping. We show that TLA can also be used to uncover insertion sites and sequences of integrated transgenes and viruses. TLA therefore promises to be a useful method in genetic research and diagnostics when comprehensive or allele-specific genetic information is needed.
journal_name
Nat Biotechnoljournal_title
Nature biotechnologyauthors
de Vree PJ,de Wit E,Yilmaz M,van de Heijning M,Klous P,Verstegen MJ,Wan Y,Teunissen H,Krijger PH,Geeven G,Eijk PP,Sie D,Ylstra B,Hulsman LO,van Dooren MF,van Zutven LJ,van den Ouweland A,Verbeek S,van Dijk KW,Corneldoi
10.1038/nbt.2959subject
Has Abstractpub_date
2014-10-01 00:00:00pages
1019-25issue
10eissn
1087-0156issn
1546-1696pii
nbt.2959journal_volume
32pub_type
杂志文章abstract：:Fluorescence microscopy of living cells enables visualization of the dynamics and interactions of intracellular molecules. However, fluorescence live-cell imaging is limited by photobleaching and phototoxicity induced by the excitation light. Here we describe controlled light-exposure microscopy (CLEM), a simple imagi...
journal_title：Nature biotechnology
pub_type： 杂志文章
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abstract：:Despite recent advances in optogenetics, it remains challenging to manipulate gene expression in specific populations of neurons. We present a dual-protein switch system, Cal-Light, that translates neuronal-activity-mediated calcium signaling into gene expression in a light-dependent manner. In cultured neurons and br...
journal_title：Nature biotechnology
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abstract：:We introduce Sailfish, a computational method for quantifying the abundance of previously annotated RNA isoforms from RNA-seq data. Because Sailfish entirely avoids mapping reads, a time-consuming step in all current methods, it provides quantification estimates much faster than do existing approaches (typically 20 ti...
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更新日期：2000-10-01 00:00:00
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journal_title：Nature biotechnology
pub_type： 杂志文章
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journal_title：Nature biotechnology
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更新日期：2003-02-01 00:00:00
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journal_title：Nature biotechnology
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journal_title：Nature biotechnology
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