Bisulphite Conversion
Bisulphite Conversion - Related Videos
04:36
Bisulfite Conversion of Genomic DNA: A Method to Study DNA Methylation in Genomic DNA Samples from Gastrointestinal Cancer Tissue
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2023
This video demonstrates the procedure of bisulfite modification of genomic DNA from gastrointestinal cancer samples. The technique is beneficial for investigating the relationship between genes’ methylation patterns and establishing their contribution to carcinogenesis.
13:18
Single Oocyte Bisulfite Mutagenesis
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Cited by 14 •
2012
Bisulfite mutagenesis is the gold standard for analyzing DNA methylation. Our modified protocol allows for DNA methylation analysis at the single-cell level and was specifically designed for individual oocytes. It can also be used for cleavage-stage embryos.
02:08
Gene Conversion
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2020
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
02:08
Gene Conversion
0 Views •
2023
Other than maintaining genome stability via DNA repair, homologous recombination plays an important role in diversifying the genome. In fact, the recombination of sequences forms the molecular basis of genomic evolution. Random and non-random permutations of genomic sequences create a library of new amalgamated sequences. These newly formed genomes can determine the fitness and survival of cells. In bacteria, homologous and non-homologous types of recombination lead to the evolution of new...
01:36
Conversion of Units
0 Views •
2023
Sometimes, there is a need to convert from one unit to another one. For instance, reading a cookbook in which quantities are expressed in units of liters or ounces may require conversion of quantities to cups. Or, when looking up directions on how to get to a location, we may be interested to know how many miles we are going to walk. In this case, we would have to convert units of feet or meters to miles. The first step in the unit conversion is to list the given units and the units required...
FAQs
The key analytical signal comes from different chemical fates of cytosines during treatment. Unmethylated cytosines are converted to uracil, whereas 5-methylcytosines are largely protected. Following amplification, uracil is represented as thymine and protected methylated positions remain read as cytosine, allowing sequence data to distinguish the two original cytosine states.
Treatment alone does not provide the final methylation readout. Amplification incorporates the conversion differences into DNA that can be examined, and sequencing then records thymine or cytosine signals at the relevant positions. Comparing those signals across a target region makes it possible to map methylation patterns rather than merely detect that DNA underwent chemical treatment.
Converted positions provide a pattern of methylated and unmethylated cytosines across a DNA region. Examining that pattern can help researchers relate methylation states to gene regulation and other epigenetic processes. Because the method can be applied across genes or genomes, it supports analysis of localized patterns as well as broader methylation landscapes.
A typical workflow begins with DNA treatment using bisulphite, followed by amplification of the treated material. The amplified DNA is then analyzed by sequencing or by a methylation-specific assay. Researchers interpret cytosine and thymine signals at selected positions to infer the underlying methylation pattern and connect it with the biological question under study.
The choice depends on the desired scope of the analysis. Bisulphite sequencing is suited to mapping methylation patterns across genes or genomes, while methylation-specific assays provide an approach for investigating selected methylation states. Both use the conversion-dependent difference between thymine and cytosine signals, but they support different levels of regional or genome-wide analysis.
Biologists use the resulting methylation maps to investigate how epigenetic patterns relate to gene regulation, development, and disease-associated changes. The same approach can also examine other biological processes in which DNA methylation is relevant. Comparing patterns among genes, genomes, or biological conditions helps researchers identify methylation changes associated with those contexts.