Table 1
Comparison of assay conditions and composition of replicating and nonreplicating media for the
Mycobacterium tuberculosis wild-type and ΔpanCDΔlysA strains
Replicating
medium for
M. tuberculosis
wild-type
Replicating medium
for M. tuberculosis
ΔpanCDΔlysA
Non-replicating
medium for
M. tuberculosis wildtype
Non-replicating
medium for
M. tuberculosis
ΔpanCDΔlysA
Starting
inoculum
0.01
0.01
0.1
0.1
Time of
exposure to
drug for
HTS (days)
7 days
7 days
3 days in
non-replicating
conditions;
7–10 days at 1/5 or
1/21 initial
concentration in
replicating medium
during outgrowth
3 days in
non-replicating
conditions;
7–10 days at 1/5
or 1/21 initial
concentration in
replicating
medium during
outgrowth
Fold dilution
–
–
1 to 5 (384-well
format) or 1 to
21 (96-well format)
1 to 5 (384-well
format)
or 1 to 21 (96-well
format)
Time of
replicating
outgrowth
prior to
determining
OD580
–
–
7–10 days OG
7–10 days OG
Type of base
medium
Middlebrook
7H9
Middlebrook 7H9
Sauton’s
Sauton’s
pH
6.6
6.6
5.0
5.0
Carbon source Glycerol (0.2%);
dextrose (0.2%)
Glycerol (0.5%);
dextrose (0.2%)
Butyrate (0.05%)
Butyrate (0.05%)
Nitrogen
source
Glutamate
Glutamate; CAS
amino acids
(0.5%)
NH 4 Cl and ferric
ammonium citrate
NH 4 Cl and ferric
ammonium
citrate
O 2
20%
20%
1%
1%
CO 2
5%
5%
5%
5%
Source of nitric
oxide and
reactive
nitrogen
intermediates
–
–
0.5 mM NaNO 2
0.5 mM NaNO 2
(continued)
614
Ben Gold et al.