Dna Probe Preparation
Dna Probe Preparation - Related Videos
14:57
DNA Stable-Isotope Probing (DNA-SIP)
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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.
12:16
Robust 3D DNA FISH Using Directly Labeled Probes
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We describe a robust and versatile protocol for analyzing nuclear architecture by 3D DNA FISH using directly labeled fluorescent probes.
03:31
Labeling DNA Probes
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2021
DNA probes are fragments of DNA labeled with a reporter tag to enable their detection or purification. The resulting labeled DNA probes can then hybridize to target nucleic acid sequences through complementary base-pairing, and may be used to recover or identify these regions. Radioisotopes, fluorophores, or small molecule binding partners like biotin or digoxigenin, are the most widely used reporter tags for labeling DNA probes. These labels can be attached to the probe DNA molecule via...
12:45
Preparation of Viral DNA from Nucleocapsids
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Cited by 40 •
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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.
05:58
Using DNA-Based Tension Probes to Assess Receptor Forces Applied by Immune Cells
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The video demonstrates a method for assessing receptor forces applied by immune cells using polyethylene glycol-anchored gold nanoparticles and DNA hairpin tension probes. CD8-positive T cell interactions trigger probe unfolding, emitting fluorescence while locking strands sustain the signal for quantifying the forces applied by immune cells.
FAQs
Choosing the appropriate target fragment determines which complementary nucleic acid sequence the probe can recognize. The target may be selected directly or amplified before labeling, allowing preparation to focus on a defined genetic region. This choice is important because the resulting probe connects a particular sequence with a detectable signal, supporting precise identification in downstream hybridization assays.
Enzymatic and chemical methods provide alternative ways to incorporate a detectable label into the selected DNA fragment. The overview identifies both approaches but does not specify that one is universally superior. Their shared purpose is to make sequence-specific binding observable after hybridization, so the labeling strategy should remain compatible with the intended detection assay and target application.
Purification removes unwanted components from the prepared material, while denaturation separates the probe strands before hybridization. This strand separation promotes access to complementary bases on the target nucleic acid. Together, these steps help the labeled fragment participate more effectively in sequence-specific pairing and improve the reliability of the signal produced during detection.
A typical workflow begins by selecting or amplifying the target DNA fragment, followed by incorporating a detectable label through an enzymatic or chemical method. The product is then purified and denatured before use. These stages convert sequence information into a probe suitable for complementary base pairing, while preparation quality influences how reliably the target becomes detectable.
Prepared probes support different assay formats that reveal different kinds of genetic information. In Southern blotting, they help detect specific DNA sequences, whereas fluorescence in situ hybridization uses probes to examine sequence location in relation to chromosomes. Thus, the same preparation principles can support both sequence identification and spatial analysis of genomic material.
In genetics, probe-based assays can identify specific genes and variants, locate sequences, and detect genomic rearrangements. Fluorescence in situ hybridization is especially relevant when sequence position must be considered alongside chromosome structure, while Southern blotting supports detection of defined DNA sequences. These applications link molecular sequence information with larger-scale genomic organization and function.