Molecular Biology

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    Protocols in Current Issue
    Using indCAPS to Detect CRISPR/Cas9 Induced Mutations
    Authors:  Charles Hodgens, Zachary L. Nimchuk and Joseph J. Kieber, date: 09/20/2019, view: 260, Q&A: 0
    [Abstract] Cleaved amplified polymorphic sequences (CAPS) assays are useful tools for detecting small mutations such as single nucleotide polymorphisms (SNPs) or insertion/deletions (indels) present in an amplified DNA fragment. A mutation that disrupts or creates a restriction site will prevent cleavage by a restriction enzyme, allowing discrimination of ...
    Genotyping of the OATP1B1 c. 521 T>C Polymorphism from the Formalin-Fixed Paraffin-Embedded (FFPE) Tissue Specimens: An Optimized Protocol
    Authors:  Alexandra R Crowe, Jonathan Miller and Wei Yue, date: 08/20/2019, view: 306, Q&A: 0
    [Abstract] Organic anion transporting polypeptide (OATP) 1B1 is a liver-specific transport protein that plays an important role in hepatic drug disposition. It transports many drugs from the blood into the liver, including lipid-lowering statins. The c. 521 T>C polymorphism of OATP1B1 has reduced transport activity and is associated with statin-induced ...
    A High-throughput qPCR-based Method to Genotype the SOD1G93A Mouse Model for Relative Copy Number
    Authors:  Valerie R. Tassinari and Fernando G. Vieira, date: 06/20/2019, view: 1095, Q&A: 0
    [Abstract] The most commonly used mouse model in ALS preclinical research expresses multiple copies of the human SOD1 (G93A) transgene. During the course of breeding, successive generations of mice can lose copies of the transgene. Because shorter lifespan of these mice is dependent on transgene copy number, it is essential to ensure that no low-copy, and ...
    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: 3107, 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: 4314, 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: 5588, 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: 5642, 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: 5457, 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 ...



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