Nucleic Acid Sequence
Nucleic acid sequence is determination of nucleotide bases sequence in a RNA or DNA fragment. This type of sequencing is an indispensable tool for carrying out basic biological research and it is also applied in varying fields like molecular diagnostics, forensics, systems biology, pharmacogenomics and genetic engineering.
By principle sequences are presented from the five inch to the three inch end. Normally, nucleic acids are linear (unbranched) polymers specification of the sequence is equivalent to definition of covalent structure of the complete molecule. Precisely for this reason, sequence and nucleic acid is also termed as the primary structure.
Sequence also has the capacity of representing information. Biological DNA also represents the information that is needed to direct functions of a living thing and in such context, the term genetic sequence is used often. Sequences can also be read from biological raw material though DNA sequencing techniques.
Nucleic acids also have a tertiary and secondary structure. The primary structure of often mistakenly known as the primary sequence. Equally, there is no parallel concept of tertiary and secondary sequence. The analysis of DNA sequence was traditionally accomplished through use of two varying techniques:
- The ddNTP mediated chain termination technique if Sanger (1997
- The chemical degradation technique of Gilbert and Maxam (1997).
Numerous next generation sequencing technologies for DNA have been developed and while this is the case, the traditional nucleic sequence techniques are routinely used in majority of life science laboratories. Obtaining optimal results with the conventional sequencing techniques requires careful reagents selection.
Nucleic acids is comprised of a chain of units that are linked known as nucleotides. Each nucleotide is made of 3 subunits which include, a sugar (ribose in RNA case and in DNA case deoxyribose) which makes up the backbone of nucleic acid strand, phosphate group and a set of nucleobases attached to the sugar. The nucleobases are especially important in the base pairing of strands to form tertiary and secondary structure like the famed double helix.
Possible letters in this case include A, C, G and T which represent the 4 nucelotide bases of DNA strand adenine, cytosine, guanine, thymine-covalently linked to a phosphodiester backbone. I n typical cases, the sequence are printed adjoining one another without any gaps as in the sequence AAAGTCTGAC which is read left to right in the 5 inch to 3 inch direction.
Current methods of sequencing rely on discriminatory ability of DNA polymerases and as such can only distinguish 4 bases. When using current technology, it is difficult to sequence small amounts of DNA because the signal is usually too weak t be measured and this can be overcome by polymerase chain reaction (PCR) amplification.
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