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    Protocols in Current Issue
    DNA-free Genome Editing of Chlamydomonas reinhardtii Using CRISPR and Subsequent Mutant Analysis
    Authors:  Jihyeon Yu, Kwangryul Baek, EonSeon Jin and Sangsu Bae, date: 06/05/2017, view: 3620, Q&A: 0
    [Abstract] We successfully introduced targeted knock-out of gene of interest in Chlamydomonas reinhardtii by using DNA-free CRISPR. In this protocol, the detailed procedures of an entire workflow cover from the initial target selection of CRISPR to the mutant analysis using next generation sequencing (NGS) technology. Furthermore, we introduce a ...
    Chromosome Dosage Analysis in Plants Using Whole Genome Sequencing
    Authors:  Ek Han Tan, Luca Comai and Isabelle M. Henry, date: 07/05/2016, view: 4156, Q&A: 0
    [Abstract] Relative chromosome dosage, i.e., increases or decreases in the number of copies of specific chromosome regions in one sample versus another, can be determined using aligned read-counts from Illumina sequencing (Henry et al., 2010). The following protocol was used to identify the different classes of aneuploids that ...
    Protocol for the Generation of a Transcription Factor Open Reading Frame Collection (TFome)
    [Abstract] The construction of a physical collection of open reading frames (ORFeomes) for genes of any model organism is a useful tool for the exploration of gene function, gene regulation, and protein-protein interaction. Here we describe in detail a protocol that has been used to develop the first collection of transcription factor (TF) and co-regulator ...
    Whole Genome Bisulfite Sequencing and DNA Methylation Analysis from Plant Tissue
    Authors:  Daniela Pignatta, George W. Bell and Mary Gehring, date: 02/20/2015, view: 9430, Q&A: 0
    [Abstract] This protocol describes whole genome bisulfite-sequencing library preparation from plant tissue and subsequent data analysis. Allele-specific methylation analysis and genome-wide identification of differentially methylated regions are additional features of the analysis procedure.
    Plant Sequence Capture Optimised for Illumina Sequencing
    Authors:  Axel Himmelbach, Manuela Knauft and Nils Stein, date: 07/05/2014, view: 7826, Q&A: 0
    [Abstract] Plant Sequence Capture is used for targeted resequencing of whole exomes (all exons of a genome) of complex genomes e.g. barley and its relatives (Mascher et al., 2013). Sequencing and computing costs are significantly reduced since only the greatly enriched and gene-coding part of the barley genome is targeted, that corresponds ...
    Library Construction for Genome-wide Bisulfite Sequencing in Plants
    Authors:  David Moraga Amador, Chenggang Wang, Kevin H. Holland and Zhonglin Mou, date: 12/20/2013, view: 7517, Q&A: 0
    [Abstract] DNA methylation is the most studied epigenetic modification, which involves the addition of a methyl group to the carbon-5 position of cytosine residues in DNA. DNA methylation is important for the regulation of gene expression. Bisulfite sequencing is the gold standard technique for determining genome-wide DNA methylation profiles in eukaryotes. ...