Genetic heterogeneity revealed by sequence analysis of Mycobacterium tuberculosis isolates from extra-pulmonary tuberculosis patients

Table 3 Primer sequences and standardized PCR parameters

Primer Panels	Coordinate of IS element (w.r.t. H37Rv) present in strain	Primers flanking the co-ordinate of IS element	Standardized annealing temp./ MgCl₂concentration	Amplicon size (bp)
				With IS	W/o IS
A	3480373, + strand, in AC74	5’-CAGCAGGGATGGATTCACC-3’5’-GCGAGTGGGATTCAGAGAG-3’	62°C/1mM	1836	481
B	1527976, + strand, in AC544	5’-GTGGTGCCGTCGTTGTCTC-3’5’-ATGCCCGTAATGTCTGCTGG-3’	60.2°C/1.5mM	1632	277
C	1356494, + strand, in AC741356495, + strand, in F85	5’-TTCGTCTCAATGGGCAACTAC-3’5’-CACCAGGCACTTCGTTATCG-3’	60.2°C/2.5mM	1552	197
D	2550014-2551368, + strand, in H37Rv	5’-CTGCGACTGCGTTGGTAATC-3’5’-ACGGTGGGGAAAGCCTGAAG-3’	61.3°C/1mM	1563	205
E	1541952-1543306, - strand, in H37Rv	5’-AGCAGGAGGAGCGGGACG-3’5’-GGATAACAGGCGCGAACCG-3’	72°C/1mM	1622	264
F	1657017, + strand, in F85	5’-TCCCTACACTCGGTTCATCC-3’5’-ACAGCAGCAGCGCCACGG-3’	61.3°C/1mM	1552	197
G	2229658, - strand, in F99	5’-TGGTGGTCAGGGAAAAGCC-3’5’-CCCTCCCGTAGCAGCCGC-3’	61.3°C/1.5mM	1617	262
H (IS6110^ref)	Multiple copies present in all	5’-CGTGAGGGCATCGAGGTGGC-3’5’-GCGTAGGCGTCGGTGACAAA-3’	65°C/1mM	245	0

[Ref: Hermans P.W.M., van Soolingen D., Dale J.W. et al., Insertion element IS986 from Mycobacterium tuberculosis: a Useful tool in Diagnosis and Epidemiology of tuberculosis. 1990 J. Clin. Microbiol. 28(9):2051-2058].

ISSN: 1471-2164