Molecular Biology

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    Protocols in Current Issue
    Identification of Insertion Site by RESDA-PCR in Chlamydomonas Mutants Generated by AphVIII Random Insertional Mutagenesis
    Authors:  Fantao Kong and Yonghua Li-Beisson, date: 02/05/2018, view: 2262, Q&A: 0
    [Abstract] Chlamydomonas reinhardtii is frequently used as a model organism to study fundamental processes in photosynthesis, metabolism, and flagellar biology. Versatile tool boxes have been developed for this alga (Fuhrmann et al., 1999; Schroda et al., 2000; Schroda, 2006). Among them, forward genetic approach has been ...
    Detection of Pathogens and Ampicillin-resistance Genes Using Multiplex Padlock Probes
    Authors:  Rick Conzemius and Ivan Barišić, date: 08/20/2017, view: 3704, Q&A: 0
    [Abstract] Diagnostic assays for pathogen identification and characterization are limited either by the number of simultaneously detectable targets, which rely on multiplexing methods, or by time constraints due to cultivation-based techniques. We recently presented a 100-plex method for human pathogen characterization to identify 75 bacterial and fungal ...
    Single Genome Sequencing of Expressed and Proviral HIV-1 Envelope Glycoprotein 120 (gp120) and nef Genes
    [Abstract] The current study provides detailed protocols utilized to amplify the complete HIV-1 gp120 and nef genes from single copies of expressed or integrated HIV present in fresh-frozen autopsy tissues of patients who died while on combined antiretroviral therapy (cART) with no detectable plasma viral load (pVL) at death (Lamers et al ...
    Next-generation Sequencing of the DNA Virome from Fecal Samples
    Authors:  Cynthia L. Monaco and Douglas S. Kwon, date: 03/05/2017, view: 4936, Q&A: 0
    [Abstract] Herein we describe a detailed protocol for DNA virome analysis of low input human stool samples (Monaco et al., 2016). This protocol is divided into four main steps: 1) stool samples are pulverized to evenly distribute microbial matter; 2) stool is enriched for virus-like particles and DNA is extracted by phenol-chloroform; 3) purified ...
    Generation of Mitochondrial-nuclear eXchange Mice via Pronuclear Transfer
    [Abstract] The mitochondrial paradigm for common disease proposes that mitochondrial DNA (mtDNA) sequence variation can contribute to disease susceptibility and progression. To test this concept, we developed the Mitochondrial-nuclear eXchange (MNX) model, in which isolated embryonic pronuclei from one strain of species are implanted into an enucleated ...
    EST-SSR Analysis and Cross-species Transferability Study in Lavandula
    Authors:  Ayelign M. Adal, Zerihun A. Demissie and Soheil S. Mahmoud, date: 08/05/2016, view: 4982, Q&A: 0
    [Abstract] The genus Lavandula comprises of several economically important lavender species that are mainly cultivated worldwide for essential oil production. Identification of lavender species and their cultivars has been a huge bottleneck in lavender industries due to lack of appropriate identification mechanisms. Recent advances in ...
    Identification of Natural Hybrids by SSR Markers in Mussaenda
    Authors:  Zhonglai Luo, Tingting Duan, Shuai Yuan, Shi Chen, Xiufeng Bai and Dianxiang Zhang, date: 07/05/2016, view: 4871, Q&A: 0
    [Abstract] Detection of natural hybrids is of great significance for plant taxonomy, reproductive biology, and population genetic studies. Compared with methods depending on morphological characters, molecular markers provide reliable and much more accurate results. This protocol describes approaches employing microsatellite (SSR) markers to identify ...
    Genomic DNA Extraction and Genotyping of Dictyochloropsis Green Algae Strains
    Authors:  Francesco Dal Grande, Carolina Cornejo, Christoph Scheidegger and Andreas Beck, date: 08/05/2015, view: 7250, Q&A: 0
    [Abstract] Dictyochloropsis is an ecologically important genus of free-living and symbiotic green algae. Representatives of this genus are horizontally transmitted among several fungi of the family Lobariaceae, thus forming photobiont-mediated guilds. This protocol is suitable for extracting DNA from algal cultures and lichen samples and for ...
    Pyrosequencing Approach for SNP Genotyping in Plants Using a M13 Biotinylated Primer
    Authors:  Cristina Silvar, Dragan Perovic, Anna M Casas, Ernesto Igartua and Frank Ordon, date: 05/20/2015, view: 6193, Q&A: 0
    [Abstract] Single Nucleotide Polymorphisms (SNPs), which constitute single base-pair variations in the DNA sequence, are the most abundant molecular markers in plant and animal genomes. They are becoming the markers of choice for genotyping in all fields of molecular biology, as they are easily prone to automation and high throughput, for example through ...
    Fluorescence-based CAPS Multiplex Genotyping on Capillary Electrophoresis Systems
    Authors:  Jelena Perovic, Cristina Silvar, Dragan Perovic, Nils Stein and Frank Ordon, date: 05/20/2015, view: 5764, Q&A: 0
    [Abstract] Recent advances in next-generation sequencing techniques allow the detection of a large number of SNPs and their use in a high throughput manner. However, Cleaved Amplified Polymorphic Sequences (CAPSs) still play a significant role as complement to other high throughput methods for SNP genotyping. Therefore, new methods focusing on the ...



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