Molecular Biology

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    Protocols in Current Issue
    Viral Chromosome Conformation Capture (V3C) Assays for Identifying Trans-interaction Sites between Lytic Viruses and the Cellular Genome
    Authors:  Kinjal Majumder, Maria Boftsi and David J Pintel, date: 03/20/2019, view: 14, Q&A: 0
    [Abstract] The folding mechanisms of the mammalian genome package our genetic material into the nucleus, and in doing so, dictate its appropriate replication and expression. Chromosome conformation capture technology has enabled the dissection of the folding principles of the cellular genome. This has led to a better understanding of the role played by ...
    Preparing Single-cell DNA Library Using Nextera for Detection of CNV
    Authors:  Larry Xi, Patrick Leong and Aleksandar Mihajlovic, date: 02/20/2019, view: 328, 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 ...
    Analysis of Early Phase HIV-1 Replication and Integration Events by Using Real-time PCR
    Authors:  Graf Laura, Steven P. Moran, Sally Ro, Carlos M.C. de Noronha and Binshan Shi, date: 02/20/2019, view: 327, Q&A: 0
    [Abstract] Upon entry into a host cell, the HIV-1 virus undergoes a series of critical early replication events including reverse transcription, nuclear import, and integration of its cDNA into the host genome. Molecular assays used to detect and analyze changes in HIV-1 early phase replication events are valuable tools in developing potential antiretroviral ...
    Rapid Multiplexed Reduced Representation Bisulfite Sequencing Library Prep (rRRBS)
    Authors:  Lisa-Marie Legault, Donovan Chan and Serge McGraw, date: 02/20/2019, view: 441, 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 ...
    pNP Transgenic RNAi System Manual in Drosophila
    Authors:  Fang Wang, Huan-Huan Qiao, Rong-Gang Xu, Jin Sun, Ruibao Zhu, Decai Mao and Jian-Quan Ni , date: 02/05/2019, view: 418, Q&A: 0
    [Abstract] Much of our knowledge about the mechanisms underlying biological processes relies on genetic approaches, whereby gene activity is reduced and the phenotypic consequences of perturbation are analyzed in detail. For functional genomic studies, a specific, systematic, and cost-effective manner is critical. Transgenic RNAi system is the top priority ...
    In vitro Protein-DNA Binding Assay (AlphaScreen® Technology)
    Authors:  Mika Nomoto, Yasuomi Tada and Hironaka Tsukagoshi, date: 02/05/2019, view: 635, Q&A: 0
    [Abstract] Identification of specific DNA binding sites of transcription factors is important in understanding their functions. Recent techniques allow us to investigate genome-wide in vivo binding positions by chromatin immunoprecipitation combined with high-throughput sequencing. However, to further explore the binding motifs of transcription ...
    Imaging Higher-order Chromatin Structures in Single Cells Using Stochastic Optical Reconstruction Microscopy
    Authors:  Jianquan Xu and Yang Liu, date: 02/05/2019, view: 651, Q&A: 0
    [Abstract] Higher-order chromatin organization shaped by epigenetic modifications influence the chromatin environment and subsequently regulate gene expression. Direct visualization of the higher-order chromatin structure at their epigenomic states is of great importance for understanding chromatin compaction and its subsequent effect on gene expression and ...
    Flip-flop Mediated Conditional Gene Inactivation in Drosophila
    Authors:  Sathiya N. Manivannan, Priyanka Pandey and Sonal Nagarkar-Jaiswal, date: 02/05/2019, view: 485, Q&A: 0
    [Abstract] Mosaic analysis in Drosophila, an important tool to assess cellular phenotypes of mutants in an otherwise heterozygous background, relies on mitosis. Hence, it cannot be used to inactivate gene function in mitotically inactive, terminally differentiated cells such as neurons. To address this issue, we developed “Flip-flop”, a novel, ...
    Measuring Homologous Recombination Rates between Chromosomal Locations in Salmonella
    Authors:  Gerrit Brandis, Sha Cao and Diarmaid Hughes, date: 02/05/2019, view: 407, Q&A: 0
    [Abstract] Homologous recombination between two similar DNA molecules, plays an important role in the repair of double-stranded DNA breaks. Recombination can occur between two sister chromosomes, or between two locations of similar sequence identity within the same chromosome. The assay described here is designed to measure the rate of homologous ...
    flySAM Transgenic CRISPRa System Manual
    Authors:  Yu Jia, Da Shen, Xia Wang, Jin Sun, Ping Peng, Rong-Gang Xu, Bowen Xu and Jian-Quan Ni , date: 01/20/2019, view: 877, Q&A: 0
    [Abstract] Powerful and general methods that can enhance gene expression are useful to systematically study gene function. To date, compared with the methods in generating loss-of-function mutants, methods to achieve gain-of-function are limited. The entire field in Drosophila has relied heavily on the Gal4/UAS:cDNA overexpression system developed ...



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