Molecular Biology


    Protocols in Current Issue
    Modeling NOTCH1 driven T-cell Acute Lymphoblastic Leukemia in Mice
    Authors:  Agnieszka A. Wendorff and Adolfo A. Ferrando, date: 05/20/2020, view: 756, Q&A: 0
    [Abstract] T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive hematological malignancy that arises from transformation of T-cell primed hematopoietic progenitors. Although T-ALL is a heterogenous and molecularly complex disease, more than 65% of T-ALL patients carry activating mutations in the NOTCH1 gene. The majority of T-ALL–associated ...
    ChIP-Seq from Limited Starting Material of K562 Cells and Drosophila Neuroblasts Using Tagmentation Assisted Fragmentation Approach
    Authors:  Junaid Akhtar, Piyush More and Steffen Albrecht, date: 02/20/2020, view: 722, Q&A: 0
    [Abstract] Chromatin immunoprecipitation is extensively used to investigate the epigenetic profile and transcription factor binding sites in the genome. However, when the starting material is limited, the conventional ChIP-Seq approach cannot be implemented. This protocol describes a method that can be used to generate the chromatin profiles from as low as ...
    Strand-specific Single-stranded DNA Sequencing (4S-seq) of E. coli genomes
    Authors:  Takahiro Masuda, Nobuaki Kono, Masaru Tomita and Kazuharu Arakawa, date: 08/05/2019, view: 1877, Q&A: 0
    [Abstract] Most bacterial genomes have biased nucleotide composition, and the asymmetry is considered to be caused by a single-stranded DNA (ssDNA) deamination arising from the bacterial replication machinery. In order to evaluate the relationship experimentally, the position and frequency of ssDNA formed during replication must be verified clearly. Although ...
    Detection of Heteroplasmic Variants in the Mitochondrial Genome through Massive Parallel Sequencing
    Authors:  Joke Mertens, Filippo Zambelli, Dorien Daneels, Ben Caljon, Karen Sermon and Claudia Spits, date: 07/05/2019, view: 1814, Q&A: 0
    [Abstract] Detecting heteroplasmies in the mitochondrial DNA (mtDNA) has been a challenge for many years. In the past, Sanger sequencing was the main option to perform this analysis, however, this method could not detect low frequency heteroplasmies. Massive Parallel Sequencing (MPS) provides the opportunity to study the mtDNA in depth, but a controlled ...
    Improved HTGTS for CRISPR/Cas9 Off-target Detection
    Authors:  Jianhang Yin, Mengzhu Liu, Yang Liu and Jiazhi Hu, date: 05/05/2019, view: 2298, Q&A: 0
    [Abstract] Precise genome editing is essential for scientific research and clinical application. At present, linear amplification-mediated high-throughput genome-wide translocation sequencing (LAM-HTGTS) is one of most effective methods to evaluate the off-target activity of CRISPR-Cas9, which is based on chromosomal translocation and employs a “bait” DNA ...
    Preparing Single-cell DNA Library Using Nextera for Detection of CNV
    Authors:  Larry Xi, Patrick Leong and Aleksandar Mihajlovic, date: 02/20/2019, view: 2474, Q&A: 0
    [Abstract] Single-cell DNA sequencing is a powerful tool to evaluate the state of heterogeneity of heterogeneous tissues like cancer in a quantitative manner that bulk sequencing can never achieve. DOP-PCR (Degenerate Oligonucleotide-Primed Polymerase Chain Reaction), MDA (Multiple Displacement Amplification), MALBAC (Multiple Annealing and Looping-Based ...
    Rapid Multiplexed Reduced Representation Bisulfite Sequencing Library Prep (rRRBS)
    Authors:  Lisa-Marie Legault, Donovan Chan and Serge McGraw, date: 02/20/2019, view: 2586, Q&A: 0
    [Abstract] DNA methylation is a common mechanism of epigenetic regulation involved in transcriptional modulation and genome stability. With the evolution of next-generation sequencing technologies, establishing quantitative genome-wide DNA methylation profiles is becoming routine in many laboratories. However, many of these approaches take several days to ...
    Quantitative ChIP-seq by Adding Spike-in from Another Species
    Authors:  Kongyan Niu, Rui Liu and Nan Liu, date: 08/20/2018, view: 15018, Q&A: 0
    [Abstract] Chromatin immunoprecipitation followed by sequencing (ChIP-seq) is a routine procedure in the lab; however, epigenome-wide quantitative comparison among independent ChIP-seq experiments remains a challenge. Here, we contribute an experimental protocol combined with a computational workflow allowing quantitative and comparative assessment of ...
    Reduced Representation Bisulfite Sequencing in Maize
    Authors:  Fei-Man Hsu, Chung-Ju Rachel Wang and Pao-Yang Chen, date: 03/20/2018, view: 4271, Q&A: 0
    [Abstract] DNA methylation is an epigenetic modification that regulates plant development (Law and Jacobsen, 2010). Whole genome bisulfite sequencing (WGBS) is a state-of-the-art method for profiling genome-wide methylation patterns with single-base resolution (Cokus et al., 2008). However, for an organism with a large genome, e.g., the 2.1 ...

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