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Chromatin Interaction Analysis with Paired-End Tag Sequencing (ChIA-PET) for Mapping Chromatin Interactions and Understanding Transcription Regulation


JoVE 3770 4/30/2012

1Genome Institute of Singapore, Agency for Science, Technology and Research, Singapore, 2A*STAR-Duke-NUS Neuroscience Research Partnership, Singapore, 3Department of Biochemistry, National University of Singapore, Singapore

Chromatin Interaction Analysis by Paired-End Tag Sequencing (ChIA-PET) is a method for de novo detection of chromatin interactions, for better understanding of transcriptional control.

 

DNA Fingerprinting of Mycobacterium leprae Strains Using Variable Number Tandem Repeat (VNTR) - Fragment Length Analysis (FLA)


JoVE 3104 7/15/2011

Department of Microbiology, Immunology and Pathology, Colorado State University

Leprosy, caused by Mycobacterium leprae, is still endemic in many places. In order to learn about the spread and mode of transmission of leprosy, it is important to determine which strain of M. leprae has infected a patient. Variable numbers of tandem repeats (VNTR) typing is one such method.

 

Agarose Gel Electrophoresis for the Separation of DNA Fragments


JoVE 3923 4/20/2012

Department of Molecular, Cell, and Developmental Biology, University of California Los Angeles

A basic protocol for the separation of DNA fragments using agarose gel electrophoresis is described.

 

Isolation of Translating Ribosomes Containing Peptidyl-tRNAs for Functional and Structural Analyses


JoVE 2498 2/25/2011

1Department of Biological Sciences, University of Alabama Huntsville, 2Department of Biology, Stanford University

A major impediment to biochemical analyses of ribosomes containing nascent peptidyl-tRNAs has been the presence of other ribosomes in the same samples, ribosomes not involved in the translation of the specific mRNA sequence being analyzed. We developed a simple methodology to purify, exclusively, the ribosomes containing the nascent peptidyl-tRNA of interest.

 

Quantifying the Activity of cis-Regulatory Elements in the Mouse Retina by Explant Electroporation


JoVE 2821 6/28/2011

Department of Pathology and Immunology, Washington University School of Medicine

This protocol describes a simple and inexpensive way to quantify the activity of cis-regulatory elements (i.e., enhancer/promoters) in living mouse retinas via explant electroporation. DNA preparation, retinal dissection, electroporation, retinal explant culture, and post-fixation analysis and quantification are described.

 

DNA Methylation: Bisulphite Modification and Analysis


JoVE 3170 10/21/2011

1Epigenetics Group, Cancer Research Program, Garvan Institute of Medical Research, 2St Vincent's Clinical School, University of NSW

The gold standard for DNA methylation analysis is genomic sequencing of bisulphite converted DNA. This method takes advantage of the increased sensitivity of cytosine compared with 5-methylcytosine (5-MeC) to bisulphite deamination under acidic conditions. Unmethylated cytosines can be distinguished from methylated cytosines after PCR amplification of the target genomic DNA.

 

Streamlined Purification of Plasmid DNA From Prokaryotic Cultures


JoVE 2407 1/05/2011

Pall Life Sciences

This protocol is a cost effective alternative for efficient parallel clarification and plasmid DNA purification from E. coli cultures. The AcroPrep Advance process starts with an optimized lysate clarification filter plate followed by purification on a high binding capacity DNA binding filter plate.

 

DNA Extraction from 0.22 μM Sterivex Filters and Cesium Chloride Density Gradient Centrifugation


JoVE 1352 9/18/2009

Department of Microbiology and Immunology, University of British Columbia - UBC

We describe a method for extraction of high molecular weight genomic DNA from planktonic biomass concentrated on 0.22 μm Sterivex filters, followed by cesium chloride density gradient centrifugation for purification.

 

Extraction of High Molecular Weight DNA from Microbial Mats


JoVE 2887 7/07/2011

Department of Environmental Health Sciences, Arnold School of Public Health, University of South Carolina

We provide an improved protocol for extracting high molecular weight DNA from hypersaline microbial mats. Microbial cells are separated from the mat matrix prior to DNA extraction and purification. This enhances the concentrations, quality, and size of the DNA. The protocol may be used for other refractory samples.

 

DNA Stable-Isotope Probing (DNA-SIP)


JoVE 2027 8/02/2010

Department of Biology, University of Waterloo

DNA stable-isotope probing is a cultivation-independent method to identify and characterize active communities of microorganisms that are capable of utilizing specific substrates. Assimilation of substrate enriched in heavy isotope leads to incorporation of labelled atoms into microbial biomass. Density gradient ultracentrifugation retrieves labelled DNA for downstream molecular analyses.

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