Welcome guest, Sign in

Home

Viruses

Determination of Adeno-associated Virus Rep DNA Binding Using Fluorescence Anisotropy

Featured protocol,  Authors: Francisco Zarate-Perez
Francisco Zarate-PerezAffiliation: Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond VA, USA
Bio-protocol author page: a4284
Vishaka Santosh
Vishaka SantoshAffiliation: Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond VA, USA
Bio-protocol author page: a4286
Martino Bardelli
Martino BardelliAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4283
Leticia Agundez
Leticia AgundezAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4285
R. Michael Linden
R. Michael LindenAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4287
Els Henckaerts
Els HenckaertsAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4289
 and Carlos R. Escalante
Carlos R. EscalanteAffiliation: Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond VA, USA
For correspondence: carlos.escalante@vcuhealth.org
Bio-protocol author page: a4288
date: 3/20/2017, 93 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2194.

Brief version appeared in J Virol, Jul 2016
Quantitative measurement of proteins binding to DNA is a requisite to fully characterize the structural determinants of complex formation necessary to understand the DNA transactions that regulate cellular processes. Here we describe a detailed protocol to measure binding affinity of the adeno-associated virus (AAV) Rep68 protein for the integration site AAVS1 using fluorescent anisotropy. This protocol can be used to measure the binding constants of any DNA binding protein provided the substrate DNA is fluorescently labeled.

In vitro Assay to Assess Efficacy of Potential Antiviral Compounds against Enterovirus D68

Featured protocol,  Authors: Liang Sun
Liang SunAffiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
Bio-protocol author page: a4263
Leen Delang
Leen DelangAffiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
Bio-protocol author page: a4264
Carmen Mirabelli
Carmen Mirabelli Affiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
Bio-protocol author page: a4265
 and Johan Neyts
Johan NeytsAffiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
For correspondence: johan.neyts@kuleuven.be
Bio-protocol author page: a1546
date: 3/20/2017, 85 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2183.

Brief version appeared in Antimicrob Agents Chemother, Dec 2015
In 2014 enterovirus D68 (EV-D68) caused the largest outbreak in the United States since the discovery of the virus. Distinct from before, the 2014 infections were associated with more severe respiratory disease and occasional neurological complications. So far, there are no available vaccines or antivirals for the prophylaxis or treatment of EV-D68 infections. In order to evaluate the antiviral activity of potential inhibitors of EV-D68 replication, a cell-based cytopathic effect (CPE) reduction assay was developed (Sun et al., 2015).

Next-generation Sequencing of the DNA Virome from Fecal Samples

Featured protocol,  Authors: Cynthia L. Monaco
Cynthia L. MonacoAffiliation: Department of Medicine, Washington University School of Medicine, St. Louis, USA
For correspondence: cmonaco@wustl.edu
Bio-protocol author page: a4038
 and Douglas S. Kwon
Douglas S. KwonAffiliation 1: Ragon Institute of MGH, MIT, and Harvard, Cambridge, USA
Affiliation 2: Division of Infectious Diseases, Massachusetts General Hospital and Harvard Medical School, Boston, USA
Bio-protocol author page: a4039
date: 3/5/2017, 199 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2159.

Brief version appeared in Cell Host Microbe, Mar 2016
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 DNA is multiple-strand displacement amplified (MDA) and fragmented; and 4) libraries are constructed and sequenced using Illumina Miseq. Subsequent sequence analysis for viral sequence identification should be sensitive but stringent.

Transient Transfection-based Fusion Assay for Viral Proteins

Featured protocol,  Authors: Melina Vallbracht
Melina VallbrachtAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
Bio-protocol author page: a4197
Christina Schröter
Christina SchröterAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
Bio-protocol author page: a4205
Barbara G. Klupp
Barbara G. KluppAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
Bio-protocol author page: a4206
 and Thomas C. Mettenleiter
Thomas C. MettenleiterAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
For correspondence: thomas.mettenleiter@fli.de
Bio-protocol author page: a4207
date: 3/5/2017, 178 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2162.

Brief version appeared in J Virol, Dec 2015
Membrane fusion is vital for entry of enveloped viruses into host cells as well as for direct viral cell-to-cell spread. To understand the fusion mechanism in more detail, we use an infection free system whereby fusion can be induced by a minimal set of the alphaherpesvirus pseudorabies virus (PrV) glycoproteins gB, gH and gL. Here, we describe an optimized protocol of a transient transfection based fusion assay to quantify cell-cell fusion induced by the PrV glycoproteins.

Determination of Adeno-associated Virus Rep DNA Binding Using Fluorescence Anisotropy

Authors: Francisco Zarate-Perez
Francisco Zarate-PerezAffiliation: Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond VA, USA
Bio-protocol author page: a4284
Vishaka Santosh
Vishaka SantoshAffiliation: Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond VA, USA
Bio-protocol author page: a4286
Martino Bardelli
Martino BardelliAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4283
Leticia Agundez
Leticia AgundezAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4285
R. Michael Linden
R. Michael LindenAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4287
Els Henckaerts
Els HenckaertsAffiliation: Department of Infectious Diseases, King’s College London, London, United Kingdom
Bio-protocol author page: a4289
 and Carlos R. Escalante
Carlos R. EscalanteAffiliation: Department of Physiology and Biophysics, Virginia Commonwealth University School of Medicine, Richmond VA, USA
For correspondence: carlos.escalante@vcuhealth.org
Bio-protocol author page: a4288
date: 3/20/2017, 93 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2194.

[Abstract] Quantitative measurement of proteins binding to DNA is a requisite to fully characterize the structural determinants of complex formation necessary to understand the DNA transactions that regulate cellular processes. Here we describe a detailed protocol to measure binding affinity of the adeno-associated ...

In vitro Assay to Assess Efficacy of Potential Antiviral Compounds against Enterovirus D68

Authors: Liang Sun
Liang SunAffiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
Bio-protocol author page: a4263
Leen Delang
Leen DelangAffiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
Bio-protocol author page: a4264
Carmen Mirabelli
Carmen Mirabelli Affiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
Bio-protocol author page: a4265
 and Johan Neyts
Johan NeytsAffiliation: KU Leuven – University of Leuven, Department of Microbiology and Immunology, Rega Institute for Medical Research, Laboratory of Virology and Chemotherapy, Leuven, Belgium
For correspondence: johan.neyts@kuleuven.be
Bio-protocol author page: a1546
date: 3/20/2017, 85 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2183.

[Abstract] In 2014 enterovirus D68 (EV-D68) caused the largest outbreak in the United States since the discovery of the virus. Distinct from before, the 2014 infections were associated with more severe respiratory disease and occasional neurological complications. So far, there are no available vaccines or antivirals ...

Next-generation Sequencing of the DNA Virome from Fecal Samples

Authors: Cynthia L. Monaco
Cynthia L. MonacoAffiliation: Department of Medicine, Washington University School of Medicine, St. Louis, USA
For correspondence: cmonaco@wustl.edu
Bio-protocol author page: a4038
 and Douglas S. Kwon
Douglas S. KwonAffiliation 1: Ragon Institute of MGH, MIT, and Harvard, Cambridge, USA
Affiliation 2: Division of Infectious Diseases, Massachusetts General Hospital and Harvard Medical School, Boston, USA
Bio-protocol author page: a4039
date: 3/5/2017, 199 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2159.

[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 ...

Transient Transfection-based Fusion Assay for Viral Proteins

Authors: Melina Vallbracht
Melina VallbrachtAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
Bio-protocol author page: a4197
Christina Schröter
Christina SchröterAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
Bio-protocol author page: a4205
Barbara G. Klupp
Barbara G. KluppAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
Bio-protocol author page: a4206
 and Thomas C. Mettenleiter
Thomas C. MettenleiterAffiliation: Institute of Molecular Virology and Cell Biology, Friedrich-Loeffler-Institut, 17493 Greifswald-Insel Riems, Germany
For correspondence: thomas.mettenleiter@fli.de
Bio-protocol author page: a4207
date: 3/5/2017, 178 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2162.

[Abstract] Membrane fusion is vital for entry of enveloped viruses into host cells as well as for direct viral cell-to-cell spread. To understand the fusion mechanism in more detail, we use an infection free system whereby fusion can be induced by a minimal set of the alphaherpesvirus pseudorabies virus (PrV) ...

Infection of Nicotiana benthamiana Plants with Potato Virus X (PVX)

Authors: Emmanuel Aguilar
Emmanuel AguilarAffiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain
Bio-protocol author page: a3882
Francisco J. del Toro
Francisco J. del ToroAffiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain
Bio-protocol author page: a3883
Bong-Nam Chung
Bong-Nam ChungAffiliation: National Institute of Horticultural & Herbal Science, Agricultural Research Center for Climate Change, Jeju Island, South Korea
Bio-protocol author page: a3884
Tomás Canto
Tomás CantoAffiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain
Bio-protocol author page: a3885
 and Francisco Tenllado
Francisco TenlladoAffiliation: Department of Environmental Biology, Centro de Investigaciones Biológicas (CIB-CSIC), Madrid, Spain
For correspondence: tenllado@cib.csic.es
Bio-protocol author page: a3886
date: 12/20/2016, 549 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2063.

[Abstract] Potato Virus X (PVX) is the type member of Potexvirus genus, a group of plant viruses with a positive-strand RNA genome (~6.4 kb). PVX is able to establish compatible infections in Nicotiana benthamiana, a commonly used host in plant virology, leading to mild symptoms, such as chlorotic mosaic and mottling. ...

Mouse Model of Dengue Virus Infection with Serotypes 1 and 2 Clinical Isolates

Authors: Satoru Watanabe
Satoru Watanabe Affiliation: Emerging Infectious Diseases, Duke-NUS Medical School, Singapore, Singapore
For correspondence: satoru.watanabe@duke-nus.edu.sg
Bio-protocol author page: a3270
Kitti Wing Ki Chan
Kitti Wing Ki ChanAffiliation: Emerging Infectious Diseases, Duke-NUS Medical School, Singapore, Singapore
Bio-protocol author page: a3810
 and Subhash G. Vasudevan
Subhash G. VasudevanAffiliation: Emerging Infectious Diseases, Duke-NUS Medical School, Singapore, Singapore
For correspondence: subhash.vasudevan@duke-nus.edu.sg
Bio-protocol author page: a3811
date: 12/5/2016, 576 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2040.

[Abstract] Dengue is a global public health threat caused by infection with any of the 4 related dengue virus serotypes (DENV1-4). Clinical manifestations range from self-limiting febrile illness, known as dengue fever (DF), to life-threatening severe diseases, such as dengue hemorrhagic fever (DHF) or dengue ...

Quantitative Measurements of HIV-1 and Dextran Capture by Human Monocyte-derived Dendritic Cells (MDDCs)

Authors: Mickaël M. Ménager
Mickaël M. MénagerAffiliation: The Kimmel Center for Biology and Medicine of the Skirball Institute, New York University School of Medicine, New York, USA
For correspondence: mickael.menager@med.nyu.edu
Bio-protocol author page: a3703
 and Dan R. Littman
Dan R. LittmanAffiliation 1: The Kimmel Center for Biology and Medicine of the Skirball Institute, New York University School of Medicine, New York, USA
Affiliation 2: Howard Hughes Medical Institute, Washington, USA
Bio-protocol author page: a3704
date: 11/20/2016, 662 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2004.

[Abstract] The aim of this protocol is to describe how to measure and quantify the amount of HIV-1 particles and dextran molecules internalized in human monocyte derived dendritic cells (MDDCs), using three different techniques: flow cytometry, quantitative PCR and confocal microscopy....

Biochemical Analysis of Caspase-8-dependent Proteolysis of IRF3 in Virus-infected Cells

Authors: Gayatri Subramanian
Gayatri SubramanianAffiliation: Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, Toledo, USA
Bio-protocol author page: a3735
Karen Pan
Karen PanAffiliation: Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, Toledo, USA
Bio-protocol author page: a3736
Ritu Chakravarti
Ritu ChakravartiAffiliation: Department of Surgery, University of Toledo College of Medicine and Life Sciences, Toledo, USA
Bio-protocol author page: a3737
 and Saurabh Chattopadhyay
Saurabh ChattopadhyayAffiliation: Department of Medical Microbiology and Immunology, University of Toledo College of Medicine and Life Sciences, Toledo, USA
For correspondence: Saurabh.Chattopadhyay@UToledo.edu
Bio-protocol author page: a3738
date: 11/20/2016, 730 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.2018.

[Abstract] Interferon regulatory factor 3 (IRF3) is a transcription factor, which is critical for the antiviral response against a wide range of viruses (Hiscott, 2007; Ikushima et al., 2013). It gets activated in virus-infected cells via Toll like receptors (TLRs), RIG-I (retinoic acid inducible gene 1) like ...

Sequencing of Ebola Virus Genomes Using Nanopore Technology

Author: Thomas Hoenen
Thomas HoenenAffiliation: Friedrich-Loeffler-Institut, Greifswald – Isle of Riems, Germany
For correspondence: thomas.hoenen@fli.de
Bio-protocol author page: a3685
date: 11/5/2016, 741 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1998.

[Abstract] Sequencing of virus genomes during disease outbreaks can provide valuable information for diagnostics, epidemiology, and evaluation of potential countermeasures. However, particularly in remote areas logistical and technical challenges can be significant. Nanopore sequencing provides an alternative ...

Mungbean Yellow Mosaic India Virus (MYMIV)-infection, Small RNA Library Construction and Deep Sequencing for MicroRNA Identification in Vigna mungo

Authors: Anirban Kundu
Anirban KunduAffiliation: Division of Plant Biology, Bose Institute, Kolkata, West Bengal, India
Bio-protocol author page: a2047
Sujay Paul
Sujay PaulAffiliation: Division of Plant Biology, Bose Institute, Kolkata, West Bengal, India
Bio-protocol author page: a2046
Amita Pal
Amita PalAffiliation: Division of Plant Biology, Bose Institute, Kolkata, West Bengal, India
For correspondence: amita@jcbose.ac.in
Bio-protocol author page: a2048
 and Genotypic Technology
Genotypic TechnologyAffiliation: Genotypic Technology Private Limited, #2/13, Balaji Complex, Bangalore, India
Bio-protocol author page: a3589
date: 10/20/2016, 650 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1961.

[Abstract] This protocol describes small RNA library preparation from Vigna mungo total RNA followed by deep sequencing and analysis for microRNA identification.​...
1 2 3 4 5 6 7 8 9 10 

[Bio101] Lentivirus Production

Author: Nabila Aboulaich date: 3/5/2011, 22841 views, 6 Q&A
DOI: https://doi.org/10.21769/BioProtoc.39.

[Abstract] Lentivirus is a common tool used to introduce a gene into mammalian or other animal cells.This protocol is to produce lentivirus stocks from hairpin-pLKO.1 plasmid....

[Bio101] Purification of Adenovirus by Cesium Chloride Density Gradients

Author: Huan Pang
Huan PangAffiliation: Molecular Pharmacology, Albert Einstein College of Medicine, Bronx, USA
For correspondence: pang_huan@hotmail.com
Bio-protocol author page: a48
date: 4/5/2012, 17507 views, 1 Q&A
DOI: https://doi.org/10.21769/BioProtoc.201.

[Abstract] Adenovirus are efficient gene delivery systems. The standard method for purification of adenoviral vectors is based on using a cesium chloride (CsCl) density gradient combined with ultracentrifugation. This method is suitable for small-scale purification and is less expensive than column chromatography ...

Infectious Focus Assays and Multiplicity of Infection (MOI) Calculations for Alpha-herpesviruses

Authors: Anna Sloutskin
Anna SloutskinAffiliation: Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel
Bio-protocol author page: a1808
 and Ronald S. Goldstein
Ronald S. GoldsteinAffiliation: Mina and Everard Goodman Faculty of Life Sciences, Bar-Ilan University, Ramat-Gan, Israel
For correspondence: ron.goldstein@biu.ac.il
Bio-protocol author page: a1809
date: 11/20/2014, 9452 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1295.

[Abstract] Titration of viral stocks is a critical process before any experimental use of the virus. Here we describe an infectious focus assay for several alphaherpesviruses, a titration method for fluorescently labeled viruses, based on the original plaque assay. In addition, the calculation of multiplicity ...

ADCC Assay Protocol

Authors: Vikram Srivastava
Vikram SrivastavaAffiliation: Department of Microbiology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China
Bio-protocol author page: a1115
Zheng Yang
Zheng YangAffiliation: Department of Microbiology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China
Bio-protocol author page: a1116
Ivan Fan Ngai Hung
Ivan Fan Ngai HungAffiliation: Department of Medicine, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China
Bio-protocol author page: a1117
Jianqing Xu
Jianqing XuAffiliation: Institutes of Biomedical Sciences, Shanghai Public Health Clinical Center, Fudan University, Shanghai, China
Bio-protocol author page: a1118
Bojian Zheng
Bojian ZhengAffiliation: Department of Microbiology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China
Bio-protocol author page: a1119
 and Mei-Yun Zhang
Mei-Yun ZhangAffiliation: Department of Microbiology, Li Ka Shing Faculty of Medicine, University of Hong Kong, Hong Kong, China
For correspondence: zhangmy@hku.hk
Bio-protocol author page: a1120
date: 1/20/2014, 8833 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1029.

[Abstract] Antibody-dependent cell-mediated cytotoxicity (ADCC) bridges innate and adaptive immunity, and it involves both humoral and cellular immune responses. ADCC has been found to be a main route of immune protection against viral infections and cancers in vivo. Here we developed a flow cytometry based protocol ...

[Bio101] VSVG Psudotyped Retrovirus Production

Author: Jia Li
Jia LiAffiliation: Department of Immunology, Medical Center, Duke University, Durham, North Carolina, USA
For correspondence: jiali.email@gmail.com
Bio-protocol author page: a16
date: 12/5/2011, 8659 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.158.

[Abstract] Retrovirus vector pseudotyped with vesicular stomatitis virus G (VSV-G) protein has been proven to exhibit high efficiency to deliver genes in a variety of cells. Efficiency is affected by relative cell growth rate and phosphatidylserine level on the cell membrane....

Assay to Evaluate Vascular Permeability Induction in Mice

Authors: Henry Puerta-Guardo
Henry Puerta-GuardoAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a980
Arturo Raya-Sandino
Arturo Raya-SandinoAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a981
Lorenza González-Mariscal
Lorenza González-MariscalAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a982
Victor H. Rosales
Victor H. RosalesAffiliation: General Services Laboratory, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a983
José Ayala-Dávila
José Ayala-DávilaAffiliation: Department of Physiolology, Biophysics and Neurosciences, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a984
Bibiana Chávez-Mungía
Bibiana Chávez-MungíaAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a985
Daniel Martínez-Fong
Daniel Martínez-FongAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a986
Fernando Medina
Fernando MedinaAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
Bio-protocol author page: a987
Juan E. Ludert
Juan E. LudertAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
For correspondence: jludert@cinvestav.mx
Bio-protocol author page: a988
 and Rosa María del Angel
Rosa María del AngelAffiliation: Infectomics and Molecular Pathogenesis, Center for Research and Advanced Studies, Instituto Politécnico Nacional (CINVESTAV-IPN), Mexico City, México
For correspondence: rmangel@cinvestav.mx
Bio-protocol author page: a989
date: 11/20/2013, 7979 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.977.

[Abstract] Dengue virus infection usually courses as a benign self-limited fever, called dengue fever. However, on occasions it can progress to a life-threatening complication known as severe dengue (SD). A hallmark of SD is a sharp increase in vascular permeability. Secondary infections are considered a risk ...

Immunoplaque Assay (Influenza Virus)

Authors: Longping V. Tse
Longping V. TseAffiliation: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, USA
For correspondence: lt273@cornell.edu
Bio-protocol author page: a940
Yueting Zhang
Yueting ZhangAffiliation: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, USA
Bio-protocol author page: a941
 and Gary R. Whittaker
Gary R. WhittakerAffiliation: Department of Microbiology and Immunology, College of Veterinary Medicine, Cornell University, Ithaca, USA
For correspondence: grw7@cornell.edu
Bio-protocol author page: a942
date: 11/5/2013, 6334 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.959.

[Abstract] Despite developed long time ago, plaque assay is still the gold standard for viral titer quantification in modern virology. The standard crystal violet-based plaque assay relies on virus’ ability to induce cytopathic effect (CPE) which limits the assay to lytic viruses. Alternative viral quantification ...

Amplification of HIV-1 Infectious Virus in BL3 Lab

Author: Xin Wang
Xin WangAffiliation: Department of Genetics, Stanford University, Stanford, USA
For correspondence: xinwang73@hotmail.com
Bio-protocol author page: a26
date: 3/5/2012, 5865 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.102.

[Abstract] This method is used for making high titer human immunodeficiency virus type-1 (HIV-1) virus stock for subsequent infection assays. The amplification of T-tropic HIV-1 virus (IIIB strain) uses the CD4+ T cell line H9....

Packaging of Retroviral RNA into Viral Particles Analyzed by Quantitative Reverse Transcriptase-PCR

Authors: Bianca Hoffmann
Bianca HoffmannAffiliation: Department of Molecular and Medical Virology, Ruhr-University Bochum, Bochum, Germany
Bio-protocol author page: a446
 and Bastian Grewe
Bastian GreweAffiliation: Department of Molecular and Medical Virology, Ruhr-University Bochum, Bochum, Germany
For correspondence: bastian.grewe@rub.de
Bio-protocol author page: a371
date: 4/20/2013, 5513 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.684.

[Abstract] Formation of viral particles and packaging of genomic retroviral RNA into these particles are important steps in the late phase of the viral replication cycle. The efficiency of the incorporation of viral or cellular RNAs into viral particles can be studied using a quantitative Reverse Transcriptase-PCR ...

RNA Isolation and Northern Blot Analysis

Authors: Ying Liao
Ying LiaoAffiliation: School of Biological Sciences, Nanyang Technological University, Singapore, Singapore
Bio-protocol author page: a1236
To Sing Fung
To Sing FungAffiliation: School of Biological Sciences, Nanyang Technological University, Singapore, Singapore
Bio-protocol author page: a1237
Mei Huang
Mei HuangAffiliation: School of Biological Sciences, Nanyang Technological University, Singapore, Singapore
Bio-protocol author page: a1238
Shouguo Fang
Shouguo FangAffiliation: School of Biological Sciences, Nanyang Technological University, Singapore, Singapore
Bio-protocol author page: a1239
Yanxin Zhong
Yanxin ZhongAffiliation: School of Biological Sciences, Nanyang Technological University, Singapore, Singapore
Bio-protocol author page: a1240
 and Dingxiang Liu
Dingxiang LiuAffiliation: School of Biological Sciences, Nanyang Technological University, Singapore, Singapore
For correspondence: dxliu@ntu.edu.sg
Bio-protocol author page: a1241
date: 3/20/2014, 4630 views, 0 Q&A
DOI: https://doi.org/10.21769/BioProtoc.1077.

[Abstract] The northern blot is a technique used in molecular biology research to study gene expression by detection of RNA in a sample. With northern blotting it is possible to observe particular gene expression levels during differentiation, morphogenesis, as well as abnormal or diseased conditions. Here, we ...
1 2 3 4 5 6 7 8 9