What is bisulfite sequencing?
bisulfite sequencing is a method in which different areas of DNA are analyzed by methylation. Methylation is the process of adding a specific molecule called methyl group, nucleotide, in this case usually cytosine. Inactive nucleotides are often methyled, so this method can be used for different purposes, from the determination of active genome areas to identification of genes rich. In bisulfite sequencing, methylized cytosins are not affected by the sequencing process, while not methylated cytosins are converted into uracil, nucleotide, which is usually found in genetic material, deoxyribonucleic acid (bottom. Cytosin will undergo this change.vated cytosine in the genome considered.
As well as all experimental protocols, bisulfite sequencing has disadvantages. Its most important disadvantage is that for proper work it requires a very high concentration of salt. Salt promotes an annealing of a single -threaded DNA into a more natural double helix and sodium bisulfit cannot always reach cytosins when they are part of a two -fiber DNA. If the salt concentration is too high, a number of cytosins may not be converted into Uracil, resulting in a false identification of methyl cytosins in the genome. A denaturation agents may be necessary to minimize the number of false positive identification.
Great amounts genomic data are not necessary for bisulfite sequencing, so the method has a useful application analyzing clinical samples. The original source of nucleic acid does not matter, but the source must be DNA. Theoretically, it could be sequenced by this method toYselin Ribonucle (RNA), because most RNA is one -off and would not be so susceptible to false positives due to blocked nucleotides. When putting into practice, however, the sequencing of bisulfit is not useful for RNA because RNA naturally uracil. Without any kind of external marking or replenishment of the protocol, the converted cytosins would be indistinguishable from the natural uracil.
When performing any type of sequencing, accuracy and accuracy methodology, they are essential. Sensitive methods such as bisulfite sequencing offer a reliable means of sequential analysis, which in turn allows gene analysis and identification of goals for drugs and therapy. Although this method cannot be used for living people, it can still be a helpful help with the smallest tissue samples you can work with.