Dna Preparation
Dna Preparation - Related Videos
12:45
Preparation of Viral DNA from Nucleocapsids
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Cited by 40 •
2011
We describe the process of isolating high purity herpesvirus nucleocapsid DNA from infected cells. The final DNA captured from solution is of high concentration and purity, making it ideally suited for high-throughput sequencing, high fidelity PCR reactions, and transfections to produce new viral recombinants.
09:06
Preparation of DNA-crosslinked Polyacrylamide Hydrogels
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Cited by 5 •
2014
Our laboratory has developed DNA-crosslinked polyacrylamide hydrogels, a dynamic hydrogel system, to better understand the effects of modulating tissue stiffness on cell function. Here, we provide schematics, descriptions, and protocols to prepare these hydrogels.
12:03
Preparation of Mica and Silicon Substrates for DNA Origami Analysis and Experimentation
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Cited by 9 •
2015
Reproducible cleaning processes for substrates used in DNA origami research are described, including bench-top RCA cleaning and derivatization of silicon oxide. Protocols for surface preparation, DNA origami deposition, drying parameters, and simple experimental set-ups are illustrated.
08:30
Preparation of Formalin-fixed Paraffin-embedded Tissue Cores for both RNA and DNA Extraction
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Cited by 53 •
2016
This modified extraction protocol improves RNA and DNA yields from more precisely targeted regions of interest in histopathologic tissue blocks.
09:31
Amplification, Next-generation Sequencing, and Genomic DNA Mapping of Retroviral Integration Sites
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Cited by 56 •
2016
We describe a protocol for amplifying retroviral integration sites from the genomic DNA of infected cells, sequencing the amplified virus-host junctions, and then mapping these sequences to a reference genome. We also describe techniques to quantify the distribution of integration sites relative to various genomic annotations using BEDTools.
FAQs
Silica membranes and magnetic beads provide a selective capture step after cellular disruption and contaminant removal. DNA binds to the capture material, while washing helps separate remaining unwanted substances before the DNA is released into a clean solution. Choosing either approach supports recovery of DNA suitable for later amplification, sequencing, pathogen identification, or genetic analysis.
Contaminants can reduce the reliability of downstream analysis even when DNA is recovered. Proteins, lipids, RNA, and residual cellular material may interfere with assays that depend on clean DNA input. Removing these substances improves sample purity and helps produce more accurate results in polymerase chain reaction, sequencing, microbial genotyping, and related measurements.
Yield determines whether enough DNA is available for the intended analysis, while purity reflects how effectively unwanted substances were removed. Both properties contribute to reliable assay performance. A preparation with consistent yield and purity supports clearer interpretation of pathogen detection, genetic characterization, and host-response studies than a variable or contaminated sample.
A typical workflow begins by disrupting cells or other biological material to release DNA. The preparation then removes proteins, lipids, RNA, and additional contaminants, followed by DNA capture on a silica membrane or magnetic beads. Washing cleans the captured material, and elution transfers the purified DNA into a solution compatible with downstream testing.
Purified DNA can support polymerase chain reaction, sequencing, pathogen identification, microbial genotyping, and analysis of host genetic responses. These methods use the prepared material for different purposes, including detecting infectious agents, examining genetic variation, or characterizing microbial and host-associated sequences. Consistent preparation strengthens the reliability of comparisons across samples.
In infection studies, prepared DNA helps researchers identify pathogens and characterize genetic differences among microbial samples. The same general workflow can also provide material for examining host genetic responses associated with disease. This combination allows investigations to distinguish infectious-agent signals from host-related genetic information and to connect molecular findings with disease-associated changes.