Sequenciaton

DNA sequencing enables us to perform a thorough analysis of DNA

Pathogens
„
because it provides us with the most basic information of all: the
the sequence
of nucleotides. With this knowledge, for example, we can locate regulatory
and gene sequences, make comparisons between homologous genes across across
Sequenciating species and identify mutations.

„ Two methods were independently developed:

1. chemical
2. Enzimatic (Sanger’
(Sanger’s method). Sanger’
Sanger’s method became the standard
B.K.Kolita Kamal Jinadasa,
Jinadasa, because of its practicality
Post Harvest Technology Division,
NARA,
Colombo-
Colombo-15,
Sri Lanka.

„ Sanger’
Sanger’s method, is based on the use of dideoxynucleotides (ddNTP’
ddNTP’s) in addition to
Sequenciation results
the normal nucleotides (NTP’
NTP’s) found in DNA. Dideoxynucleotides are essentially the
same as nucleotides except they contain a hydrogen group on the 3’ carbon instead of a
hydroxyl group (OH). These modified nucleotides, when integrated into a sequence,
prevent the addition of further nucleotides..This occurs because a phosphodiester
bond cannot form between the dideoxynucleotide and the next incoming nucleotide,
and thus the DNA chain is terminated.

Blast
„ In bioinformatics,
bioinformatics, Basic Local Alignment Search Tool, or BLAST,
BLAST, is an
algorithm for comparing primary biological sequence information, such as the
amino-
amino-acid sequences of different proteins or the nucleotides of DNA
sequences.
sequences.
„ A BLAST search enables a researcher to compare a query sequence with a
library or database of sequences, and identify library sequences that resemble
the query sequence above a certain threshold.

„ To run, BLAST requires two sequences as input: a query sequence and a

sequence database.
„ BLAST searches for high scoring sequence alignments between the query
sequence and sequences in the database using a heuristic approach
approach that
approximates the Smith-
Smith-Waterman algorithm

1
 Blast results

Query sequence input

Search start

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