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Identification at a higher level of resolution by using variable nucleotide tandem repeat (VNTR) typing, facilitated by mycobacterial interspersed repetitive units- (MIRU-) VNTR, is desirable although this method is not currently available in Ethiopia.
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The DNA sequence was analyzed by using the standard nucleotide-nucleotide BLAST algorithm (www.ncbi.nlm.nih.gov).nih.gov
Sequences were analyzed by using the standard nucleotide-nucleotide BLAST program (National Center for Biotechnology Information, Bethesda, MD). and comparisons were carried out by using CLUSTAL W (European Bioinformatics Institute, Cambridge, UK).
Nucleotide changes between ancestral and current sequences were counted by using the "nucleotide substitution pattern" option in the "Seq.
All sequences obtained were characterized by using nucleotide blast (http://www.ncbi.nlm.nih.gov).nih.gov
We tested these by using nucleotide incorporation assays.
The nucleotide sequences obtained were compared with those Plasmodium spp. sequences available in public databases by using BLASTN (nucleotide-nucleotide) (available from http://www.ncbi.nlm.nih.gov/blast/).nih.gov/blast/
Second, SSPs were constructed for M. leprae and M. marinum by using the relevant aligned nucleotide for each of the previously identified variable synonymous sites in M. tuberculosis and M. bovis.
Pre-selective amplification was performed by using a tri-selective nucleotide (+3) at the 3'.
Bases are imaged by using reversible fluorophore terminator nucleotides.
Additionally, the same reaction conditions can be used to detect any variable nucleotide regardless of the nucleotide sequence flanking the variable site.
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