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Difco 7h9 broth
Serial monitoring outcomes of RIF stability in L-J medium and <t>7H9</t> broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
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1) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

2) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

3) Product Images from "Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis"

Article Title: Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0200145

Rv1460 truncation mutants are impaired for growth in vitro under standard culture conditions. (A–C) Growth of H37Rv (wild-type), three truncation (Δ Rv1460 stop) mutants and their complemented strains under standard conditions in 7H9 OADC. The results shown are the mean and standard deviation of three experiments.
Figure Legend Snippet: Rv1460 truncation mutants are impaired for growth in vitro under standard culture conditions. (A–C) Growth of H37Rv (wild-type), three truncation (Δ Rv1460 stop) mutants and their complemented strains under standard conditions in 7H9 OADC. The results shown are the mean and standard deviation of three experiments.

Techniques Used: In Vitro, Standard Deviation

Succinate dehydrogenase and aconitase activity is not impaired in Rv1460 truncation mutants. (A) Succinate dehydrogenase activity and (B) aconitase activity in the H37Rv (wild-type), Δ Rv1460 stop_5.20 and complemented strains cultured in 7H9 OADC. Activity was standardised relative to total protein. The results shown are the mean and standard deviation of five and three experiments respectively.
Figure Legend Snippet: Succinate dehydrogenase and aconitase activity is not impaired in Rv1460 truncation mutants. (A) Succinate dehydrogenase activity and (B) aconitase activity in the H37Rv (wild-type), Δ Rv1460 stop_5.20 and complemented strains cultured in 7H9 OADC. Activity was standardised relative to total protein. The results shown are the mean and standard deviation of five and three experiments respectively.

Techniques Used: Activity Assay, Cell Culture, Standard Deviation

4) Product Images from "Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp"

Article Title: Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp

Journal: Journal of Bacteriology

doi: 10.1128/JB.00759-13

Rel Mtb -mediated (p)ppGpp hydrolysis is required for growth in vitro and maintenance of ATP and GTP levels when (p)ppGpp is being produced. (A and B) An M. tuberculosis Δ rel Mtb strain containing an episomal vector expressing the WT TetR that induces expression of rel Mtb H80A from a cassette integrated at the attB site in the genome in the presence of ATc (Tet-Rel Mtb H80A strain, designated H80A here) and a WT M. tuberculosis strain containing an empty vector in the attB site and transformed with the same TetR-expressing plasmid (control strain). (A) Transcript levels in exponential-growth-phase cultures in liquid 7H9 media. rel Mtb H80A transcripts from the cassette at the attB site were detected in the Tet-Rel Mtb H80A strain, and transcripts from the endogenous rel Mtb gene were detected in the control strain. The first comparison shows the ratio of transcript levels in the presence compared to the absence of ATc in each strain (+ATc/−ATc), and it illustrates the induction of rel Mtb H80A in the Tet-Rel Mtb H80A strain when exposed to ATc and the unresponsiveness of the rel Mtb gene to ATc in the control strain. The second comparison shows the ratio of transcript levels in the Tet-Rel Mtb H80A strain compared to the control strain (H80A/control) under each condition to illustrate the low level of transcription of rel Mtb H80A in the absence of ATc compared to that of induced cultures. (B) M. tuberculosis Tet-Rel Mtb H80A and control strains were diluted and plated on 7H10 plates in the absence or presence of ATc. (C to G) M. smegmatis Δ rel Msm strains containing an episomal vector expressing the WT TetR that turns on expression of rel Mtb WT (Tet-Rel Mtb WT strain; designated the control) or rel Mtb H80A (Tet-Rel Mtb H80A strain; designated H80A here) from cassettes integrated at the attB site in the genome in the presence of ATc. (C and D) M. smegmatis strains were diluted and plated on 7H10 (C) and LB (D) plates in the absence or presence of ATc. (E) Graphic representation of the number of M. smegmatis CFU that grew in the presence compared to the absence of ATc when grown on LB, where all Tet-Rel Mtb H80A strain bacteria that grew in the presence of ATc on LB were suppressors and were no longer responsive to ATc treatment. Data are means ± SEM from 13 replicates. (F) ATP and GTP levels in M. smegmatis strains grown in the absence or presence of ATc for 6 h to an ODλ 600 of ∼0.6 in LB liquid media were measured by LC-MS/MS. The ratio of levels in the Tet-Rel Mtb H80A strain (H80A) to those in the Tet-Rel Mtb WT -expressing strain (control) under the same conditions is graphed. Data are the means ± SEM from 4 to 6 replicates. (E and F) The significance of differences was determined by calculating P values by Student's t tests; one asterisk indicates significance with a P value of
Figure Legend Snippet: Rel Mtb -mediated (p)ppGpp hydrolysis is required for growth in vitro and maintenance of ATP and GTP levels when (p)ppGpp is being produced. (A and B) An M. tuberculosis Δ rel Mtb strain containing an episomal vector expressing the WT TetR that induces expression of rel Mtb H80A from a cassette integrated at the attB site in the genome in the presence of ATc (Tet-Rel Mtb H80A strain, designated H80A here) and a WT M. tuberculosis strain containing an empty vector in the attB site and transformed with the same TetR-expressing plasmid (control strain). (A) Transcript levels in exponential-growth-phase cultures in liquid 7H9 media. rel Mtb H80A transcripts from the cassette at the attB site were detected in the Tet-Rel Mtb H80A strain, and transcripts from the endogenous rel Mtb gene were detected in the control strain. The first comparison shows the ratio of transcript levels in the presence compared to the absence of ATc in each strain (+ATc/−ATc), and it illustrates the induction of rel Mtb H80A in the Tet-Rel Mtb H80A strain when exposed to ATc and the unresponsiveness of the rel Mtb gene to ATc in the control strain. The second comparison shows the ratio of transcript levels in the Tet-Rel Mtb H80A strain compared to the control strain (H80A/control) under each condition to illustrate the low level of transcription of rel Mtb H80A in the absence of ATc compared to that of induced cultures. (B) M. tuberculosis Tet-Rel Mtb H80A and control strains were diluted and plated on 7H10 plates in the absence or presence of ATc. (C to G) M. smegmatis Δ rel Msm strains containing an episomal vector expressing the WT TetR that turns on expression of rel Mtb WT (Tet-Rel Mtb WT strain; designated the control) or rel Mtb H80A (Tet-Rel Mtb H80A strain; designated H80A here) from cassettes integrated at the attB site in the genome in the presence of ATc. (C and D) M. smegmatis strains were diluted and plated on 7H10 (C) and LB (D) plates in the absence or presence of ATc. (E) Graphic representation of the number of M. smegmatis CFU that grew in the presence compared to the absence of ATc when grown on LB, where all Tet-Rel Mtb H80A strain bacteria that grew in the presence of ATc on LB were suppressors and were no longer responsive to ATc treatment. Data are means ± SEM from 13 replicates. (F) ATP and GTP levels in M. smegmatis strains grown in the absence or presence of ATc for 6 h to an ODλ 600 of ∼0.6 in LB liquid media were measured by LC-MS/MS. The ratio of levels in the Tet-Rel Mtb H80A strain (H80A) to those in the Tet-Rel Mtb WT -expressing strain (control) under the same conditions is graphed. Data are the means ± SEM from 4 to 6 replicates. (E and F) The significance of differences was determined by calculating P values by Student's t tests; one asterisk indicates significance with a P value of

Techniques Used: In Vitro, Produced, Plasmid Preparation, Expressing, Transformation Assay, Liquid Chromatography with Mass Spectroscopy, Mass Spectrometry

5) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

6) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

7) Product Images from "Lansoprazole is an antituberculous prodrug targeting cytochrome bc1"

Article Title: Lansoprazole is an antituberculous prodrug targeting cytochrome bc1

Journal: Nature Communications

doi: 10.1038/ncomms8659

LPZS is a highly selective antituberculous drug with in vivo activity. ( a ) Intracellular ratio of LPZ ( m/z 370.0834, g mol −1 ) and its metabolite ( m/z 354.0884, g mol −1 ) determined by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) over a 48-h period in MRC-5 cells. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( b ) ESI–MS mass spectra in the range m/z 350–375 measured for experiments performed on the cell lysate of MRC-5 fibroblasts exposed to LPZ (extracted ion chromatograms can be found in Supplementary Fig. 4a,b ). ( c ) ESI–MS spectrum at m/z 354.0884 corresponding to the LPZS standard in methanol. ( d ) Structures of LPZ and LPZS. LPZS is missing the sulfoxide (red), which is essential for LPZ activity on the human proton pump. ( e ) LPZ/LPZS ratio determined by ESI-Q-TOF-MS over a 48-h period in 7H9 broth. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( f ) Dose–response curve of LPZS for Mtb grown in 7H9 broth (mean±s.d. of three individual experiments). ( g ) Survival of Mtb -infected MRC-5 fibroblasts was quantified at different concentrations of LPZS (mean±s.d. of three individual experiments). ( h ) Efficacy of LPZS in the mouse model of acute tuberculosis. Bacterial burden (c.f.u.) was determined in the lungs of four mice treated with the vehicle control (TPGS) or four mice treated with LPZS at 300 mg kg −1 b.i.d. given by oral gavage (mean±s.d., Student's t -test was used to compare groups).
Figure Legend Snippet: LPZS is a highly selective antituberculous drug with in vivo activity. ( a ) Intracellular ratio of LPZ ( m/z 370.0834, g mol −1 ) and its metabolite ( m/z 354.0884, g mol −1 ) determined by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) over a 48-h period in MRC-5 cells. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( b ) ESI–MS mass spectra in the range m/z 350–375 measured for experiments performed on the cell lysate of MRC-5 fibroblasts exposed to LPZ (extracted ion chromatograms can be found in Supplementary Fig. 4a,b ). ( c ) ESI–MS spectrum at m/z 354.0884 corresponding to the LPZS standard in methanol. ( d ) Structures of LPZ and LPZS. LPZS is missing the sulfoxide (red), which is essential for LPZ activity on the human proton pump. ( e ) LPZ/LPZS ratio determined by ESI-Q-TOF-MS over a 48-h period in 7H9 broth. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( f ) Dose–response curve of LPZS for Mtb grown in 7H9 broth (mean±s.d. of three individual experiments). ( g ) Survival of Mtb -infected MRC-5 fibroblasts was quantified at different concentrations of LPZS (mean±s.d. of three individual experiments). ( h ) Efficacy of LPZS in the mouse model of acute tuberculosis. Bacterial burden (c.f.u.) was determined in the lungs of four mice treated with the vehicle control (TPGS) or four mice treated with LPZS at 300 mg kg −1 b.i.d. given by oral gavage (mean±s.d., Student's t -test was used to compare groups).

Techniques Used: In Vivo, Activity Assay, Mass Spectrometry, Infection, Mouse Assay

8) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

9) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

10) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

11) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

12) Product Images from "Lansoprazole is an antituberculous prodrug targeting cytochrome bc1"

Article Title: Lansoprazole is an antituberculous prodrug targeting cytochrome bc1

Journal: Nature Communications

doi: 10.1038/ncomms8659

LPZS is a highly selective antituberculous drug with in vivo activity. ( a ) Intracellular ratio of LPZ ( m/z 370.0834, g mol −1 ) and its metabolite ( m/z 354.0884, g mol −1 ) determined by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) over a 48-h period in MRC-5 cells. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( b ) ESI–MS mass spectra in the range m/z 350–375 measured for experiments performed on the cell lysate of MRC-5 fibroblasts exposed to LPZ (extracted ion chromatograms can be found in Supplementary Fig. 4a,b ). ( c ) ESI–MS spectrum at m/z 354.0884 corresponding to the LPZS standard in methanol. ( d ) Structures of LPZ and LPZS. LPZS is missing the sulfoxide (red), which is essential for LPZ activity on the human proton pump. ( e ) LPZ/LPZS ratio determined by ESI-Q-TOF-MS over a 48-h period in 7H9 broth. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( f ) Dose–response curve of LPZS for Mtb grown in 7H9 broth (mean±s.d. of three individual experiments). ( g ) Survival of Mtb -infected MRC-5 fibroblasts was quantified at different concentrations of LPZS (mean±s.d. of three individual experiments). ( h ) Efficacy of LPZS in the mouse model of acute tuberculosis. Bacterial burden (c.f.u.) was determined in the lungs of four mice treated with the vehicle control (TPGS) or four mice treated with LPZS at 300 mg kg −1 b.i.d. given by oral gavage (mean±s.d., Student's t -test was used to compare groups).
Figure Legend Snippet: LPZS is a highly selective antituberculous drug with in vivo activity. ( a ) Intracellular ratio of LPZ ( m/z 370.0834, g mol −1 ) and its metabolite ( m/z 354.0884, g mol −1 ) determined by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) over a 48-h period in MRC-5 cells. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( b ) ESI–MS mass spectra in the range m/z 350–375 measured for experiments performed on the cell lysate of MRC-5 fibroblasts exposed to LPZ (extracted ion chromatograms can be found in Supplementary Fig. 4a,b ). ( c ) ESI–MS spectrum at m/z 354.0884 corresponding to the LPZS standard in methanol. ( d ) Structures of LPZ and LPZS. LPZS is missing the sulfoxide (red), which is essential for LPZ activity on the human proton pump. ( e ) LPZ/LPZS ratio determined by ESI-Q-TOF-MS over a 48-h period in 7H9 broth. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( f ) Dose–response curve of LPZS for Mtb grown in 7H9 broth (mean±s.d. of three individual experiments). ( g ) Survival of Mtb -infected MRC-5 fibroblasts was quantified at different concentrations of LPZS (mean±s.d. of three individual experiments). ( h ) Efficacy of LPZS in the mouse model of acute tuberculosis. Bacterial burden (c.f.u.) was determined in the lungs of four mice treated with the vehicle control (TPGS) or four mice treated with LPZS at 300 mg kg −1 b.i.d. given by oral gavage (mean±s.d., Student's t -test was used to compare groups).

Techniques Used: In Vivo, Activity Assay, Mass Spectrometry, Infection, Mouse Assay

13) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

14) Product Images from "Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis"

Article Title: Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis

Journal: PLoS ONE

doi: 10.1371/journal.pone.0200145

Rv1460 truncation mutants are impaired for growth in vitro under standard culture conditions. (A–C) Growth of H37Rv (wild-type), three truncation (Δ Rv1460 stop) mutants and their complemented strains under standard conditions in 7H9 OADC. The results shown are the mean and standard deviation of three experiments.
Figure Legend Snippet: Rv1460 truncation mutants are impaired for growth in vitro under standard culture conditions. (A–C) Growth of H37Rv (wild-type), three truncation (Δ Rv1460 stop) mutants and their complemented strains under standard conditions in 7H9 OADC. The results shown are the mean and standard deviation of three experiments.

Techniques Used: In Vitro, Standard Deviation

Succinate dehydrogenase and aconitase activity is not impaired in Rv1460 truncation mutants. (A) Succinate dehydrogenase activity and (B) aconitase activity in the H37Rv (wild-type), Δ Rv1460 stop_5.20 and complemented strains cultured in 7H9 OADC. Activity was standardised relative to total protein. The results shown are the mean and standard deviation of five and three experiments respectively.
Figure Legend Snippet: Succinate dehydrogenase and aconitase activity is not impaired in Rv1460 truncation mutants. (A) Succinate dehydrogenase activity and (B) aconitase activity in the H37Rv (wild-type), Δ Rv1460 stop_5.20 and complemented strains cultured in 7H9 OADC. Activity was standardised relative to total protein. The results shown are the mean and standard deviation of five and three experiments respectively.

Techniques Used: Activity Assay, Cell Culture, Standard Deviation

15) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

16) Product Images from "Deletion of kasB in Mycobacterium tuberculosis causes loss of acid-fastness and subclinical latent tuberculosis in immunocompetent mice"

Article Title: Deletion of kasB in Mycobacterium tuberculosis causes loss of acid-fastness and subclinical latent tuberculosis in immunocompetent mice

Journal:

doi: 10.1073/pnas.0608654104

Light microscopy of M. tuberculosis cultures grow static in 7H9 broth at 37°C for 7 days. Cultures were fixed on glass slides, and acid-fast staining was performed on the fixed smears by using the BD TB fluorescent kit-T. ( Upper ) Phase-contrast
Figure Legend Snippet: Light microscopy of M. tuberculosis cultures grow static in 7H9 broth at 37°C for 7 days. Cultures were fixed on glass slides, and acid-fast staining was performed on the fixed smears by using the BD TB fluorescent kit-T. ( Upper ) Phase-contrast

Techniques Used: Light Microscopy, Staining

17) Product Images from "Mode of Action of Clofazimine and Combination Therapy with Benzothiazinones against Mycobacterium tuberculosis"

Article Title: Mode of Action of Clofazimine and Combination Therapy with Benzothiazinones against Mycobacterium tuberculosis

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00395-15

Effect of menaquinone supplementation on clofazimine (CZM), BDQ, and isoniazid (INH) against M. tuberculosis H37Rv by REMA. Cells were grown in a normal 7H9 medium (no MK-4) or in medium supplemented with increasing concentrations of menaquinone (10, 100, and 1,000 μM MK-4). REMA results are presented as mean ± SD values of triplicates. Drug concentrations are in micrograms per milliliter.
Figure Legend Snippet: Effect of menaquinone supplementation on clofazimine (CZM), BDQ, and isoniazid (INH) against M. tuberculosis H37Rv by REMA. Cells were grown in a normal 7H9 medium (no MK-4) or in medium supplemented with increasing concentrations of menaquinone (10, 100, and 1,000 μM MK-4). REMA results are presented as mean ± SD values of triplicates. Drug concentrations are in micrograms per milliliter.

Techniques Used:

18) Product Images from "Lansoprazole is an antituberculous prodrug targeting cytochrome bc1"

Article Title: Lansoprazole is an antituberculous prodrug targeting cytochrome bc1

Journal: Nature Communications

doi: 10.1038/ncomms8659

LPZS is a highly selective antituberculous drug with in vivo activity. ( a ) Intracellular ratio of LPZ ( m/z 370.0834, g mol −1 ) and its metabolite ( m/z 354.0884, g mol −1 ) determined by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) over a 48-h period in MRC-5 cells. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( b ) ESI–MS mass spectra in the range m/z 350–375 measured for experiments performed on the cell lysate of MRC-5 fibroblasts exposed to LPZ (extracted ion chromatograms can be found in Supplementary Fig. 4a,b ). ( c ) ESI–MS spectrum at m/z 354.0884 corresponding to the LPZS standard in methanol. ( d ) Structures of LPZ and LPZS. LPZS is missing the sulfoxide (red), which is essential for LPZ activity on the human proton pump. ( e ) LPZ/LPZS ratio determined by ESI-Q-TOF-MS over a 48-h period in 7H9 broth. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( f ) Dose–response curve of LPZS for Mtb grown in 7H9 broth (mean±s.d. of three individual experiments). ( g ) Survival of Mtb -infected MRC-5 fibroblasts was quantified at different concentrations of LPZS (mean±s.d. of three individual experiments). ( h ) Efficacy of LPZS in the mouse model of acute tuberculosis. Bacterial burden (c.f.u.) was determined in the lungs of four mice treated with the vehicle control (TPGS) or four mice treated with LPZS at 300 mg kg −1 b.i.d. given by oral gavage (mean±s.d., Student's t -test was used to compare groups).
Figure Legend Snippet: LPZS is a highly selective antituberculous drug with in vivo activity. ( a ) Intracellular ratio of LPZ ( m/z 370.0834, g mol −1 ) and its metabolite ( m/z 354.0884, g mol −1 ) determined by electrospray ionization quadrupole time-of-flight mass spectrometry (ESI-Q-TOF-MS) over a 48-h period in MRC-5 cells. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( b ) ESI–MS mass spectra in the range m/z 350–375 measured for experiments performed on the cell lysate of MRC-5 fibroblasts exposed to LPZ (extracted ion chromatograms can be found in Supplementary Fig. 4a,b ). ( c ) ESI–MS spectrum at m/z 354.0884 corresponding to the LPZS standard in methanol. ( d ) Structures of LPZ and LPZS. LPZS is missing the sulfoxide (red), which is essential for LPZ activity on the human proton pump. ( e ) LPZ/LPZS ratio determined by ESI-Q-TOF-MS over a 48-h period in 7H9 broth. Representative example of three individual experiments; the complete data set can be found in Supplementary Table 2 . ( f ) Dose–response curve of LPZS for Mtb grown in 7H9 broth (mean±s.d. of three individual experiments). ( g ) Survival of Mtb -infected MRC-5 fibroblasts was quantified at different concentrations of LPZS (mean±s.d. of three individual experiments). ( h ) Efficacy of LPZS in the mouse model of acute tuberculosis. Bacterial burden (c.f.u.) was determined in the lungs of four mice treated with the vehicle control (TPGS) or four mice treated with LPZS at 300 mg kg −1 b.i.d. given by oral gavage (mean±s.d., Student's t -test was used to compare groups).

Techniques Used: In Vivo, Activity Assay, Mass Spectrometry, Infection, Mouse Assay

19) Product Images from "Bioluminescence for Assessing Drug Potency against Nonreplicating Mycobacterium tuberculosis"

Article Title: Bioluminescence for Assessing Drug Potency against Nonreplicating Mycobacterium tuberculosis

Journal: Antimicrobial Agents and Chemotherapy

doi: 10.1128/AAC.00528-15

Growth curves for M. tuberculosis 18b-Lux and evaluation of luciferase expression. (A) 18b-Lux was grown at 37°C, with shaking, in glucose- or acetate-containing 7H9 medium with 50 μg/ml STR. (B) SS18b-Lux was incubated under the same
Figure Legend Snippet: Growth curves for M. tuberculosis 18b-Lux and evaluation of luciferase expression. (A) 18b-Lux was grown at 37°C, with shaking, in glucose- or acetate-containing 7H9 medium with 50 μg/ml STR. (B) SS18b-Lux was incubated under the same

Techniques Used: Luciferase, Expressing, Incubation

20) Product Images from "Discovery of a novel dehydratase of the fatty acid synthase type II critical for ketomycolic acid biosynthesis and virulence of Mycobacterium tuberculosis"

Article Title: Discovery of a novel dehydratase of the fatty acid synthase type II critical for ketomycolic acid biosynthesis and virulence of Mycobacterium tuberculosis

Journal: Scientific Reports

doi: 10.1038/s41598-020-58967-8

Inactivation of hadD Mtb alters the bacterial fitness, biofilm and colony formation, and tolerance to low temperature. Comparison of Mtb H37Rv wt, Δ hadD and the complemented Δ hadD :: hadD Mtb strains in different phenotyping assays. All of the data are representative of at least three independent experiments. ( A ) Planktonic growth. Cultures were performed under shaking (120 rpm) at 37 °C in 7H9-based medium supplemented with 0.05% (w/v) Tween-80. Data are means and average deviations of three independent experiments. Some deviation bars are too small to be visible. ( B ) Biofilm growth at the air-liquid interface. The growth was followed for three weeks at 37 °C on 7H9-based medium. Photographs were taken after three weeks. Scale bars represent 1 cm. ( C ) Colony morphotype. Five μl culture aliquots were spotted on 7H11-based medium and grown for three weeks at 37 °C. Scale bars represent 0.5 cm. ( D ) Sensitivity to low temperature. Liquid precultures were adjusted to the same OD then serially diluted, spotted onto 7H11-based medium and incubated for four weeks at 37 °C or 30 °C.
Figure Legend Snippet: Inactivation of hadD Mtb alters the bacterial fitness, biofilm and colony formation, and tolerance to low temperature. Comparison of Mtb H37Rv wt, Δ hadD and the complemented Δ hadD :: hadD Mtb strains in different phenotyping assays. All of the data are representative of at least three independent experiments. ( A ) Planktonic growth. Cultures were performed under shaking (120 rpm) at 37 °C in 7H9-based medium supplemented with 0.05% (w/v) Tween-80. Data are means and average deviations of three independent experiments. Some deviation bars are too small to be visible. ( B ) Biofilm growth at the air-liquid interface. The growth was followed for three weeks at 37 °C on 7H9-based medium. Photographs were taken after three weeks. Scale bars represent 1 cm. ( C ) Colony morphotype. Five μl culture aliquots were spotted on 7H11-based medium and grown for three weeks at 37 °C. Scale bars represent 0.5 cm. ( D ) Sensitivity to low temperature. Liquid precultures were adjusted to the same OD then serially diluted, spotted onto 7H11-based medium and incubated for four weeks at 37 °C or 30 °C.

Techniques Used: Incubation

21) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

22) Product Images from "The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis"

Article Title: The Conserved Hypothetical Protein Rv0574c Is Required for Cell Wall Integrity, Stress Tolerance, and Virulence of Mycobacterium tuberculosis

Journal: Infection and Immunity

doi: 10.1128/IAI.02274-14

Growth profiles of mycobacterial strains in broth culture. H37Rv, ΔRv0574c, and ΔRv0574c-comp strains were inoculated at an initial optical density of 0.02 in enriched 7H9 medium. (A) OD 600 . (B) Log CFU/ml of cultures.
Figure Legend Snippet: Growth profiles of mycobacterial strains in broth culture. H37Rv, ΔRv0574c, and ΔRv0574c-comp strains were inoculated at an initial optical density of 0.02 in enriched 7H9 medium. (A) OD 600 . (B) Log CFU/ml of cultures.

Techniques Used:

23) Product Images from "Poly-L-glutamate/glutamine synthesis in the cell wall of Mycobacterium bovis is regulated in response to nitrogen availability"

Article Title: Poly-L-glutamate/glutamine synthesis in the cell wall of Mycobacterium bovis is regulated in response to nitrogen availability

Journal: BMC Microbiology

doi: 10.1186/1471-2180-13-226

Biofilm and pellicle formation under low and high nitrogen condition. A . M. bovis, wild type M. smegmatis and MSFP were grown 7H9 medium to form biofilm in low and high nitrogen medium. B . Biofilm formation assayed using the 1% crystal violet (CV) staining assay. Cells in low nitrogen (black bars), High nitrogen (crossed bars) and control (grey bars) in 7H9 media were grown in low and high nitrogen on polystyrene plates. The experiments were repeated three times with similar result. Control, medium only. C . Pellicle formation at the air-liquid interface of the standing 7H9 culture by strains M. bovis (i) , M. smegmatis (ii) and MSFP (iii) in low and high nitrogen condition. Results are representative of at least three independent experiments. LN, low nitrogen; HN, high nitrogen.
Figure Legend Snippet: Biofilm and pellicle formation under low and high nitrogen condition. A . M. bovis, wild type M. smegmatis and MSFP were grown 7H9 medium to form biofilm in low and high nitrogen medium. B . Biofilm formation assayed using the 1% crystal violet (CV) staining assay. Cells in low nitrogen (black bars), High nitrogen (crossed bars) and control (grey bars) in 7H9 media were grown in low and high nitrogen on polystyrene plates. The experiments were repeated three times with similar result. Control, medium only. C . Pellicle formation at the air-liquid interface of the standing 7H9 culture by strains M. bovis (i) , M. smegmatis (ii) and MSFP (iii) in low and high nitrogen condition. Results are representative of at least three independent experiments. LN, low nitrogen; HN, high nitrogen.

Techniques Used: Staining

Growth of the mycobacterial strains in low and high nitrogen broth culture. A . OD 600 of wild type M. bovis was inoculated to an initial optical density of 0.006 - 0.008 in 7H9 medium containing (●) low nitrogen (3.8 mM ammonium sulphate) and (▲) high nitrogen (60 mM ammonium sulphate). B . OD 600 of wild type M. smegmatis and MSFP in low and high nitrogen broth culture. Wild type M. smegmatis, low nitrogen (■), high nitrogen (□); MSFP, low nitrogen (●), high nitrogen (○). Data is mean ± SD of values obtained from three independent cultures. LN, low nitrogen; HN, high nitrogen.
Figure Legend Snippet: Growth of the mycobacterial strains in low and high nitrogen broth culture. A . OD 600 of wild type M. bovis was inoculated to an initial optical density of 0.006 - 0.008 in 7H9 medium containing (●) low nitrogen (3.8 mM ammonium sulphate) and (▲) high nitrogen (60 mM ammonium sulphate). B . OD 600 of wild type M. smegmatis and MSFP in low and high nitrogen broth culture. Wild type M. smegmatis, low nitrogen (■), high nitrogen (□); MSFP, low nitrogen (●), high nitrogen (○). Data is mean ± SD of values obtained from three independent cultures. LN, low nitrogen; HN, high nitrogen.

Techniques Used:

24) Product Images from "The Rip1 Protease of Mycobacterium tuberculosis Controls the SigD Regulon"

Article Title: The Rip1 Protease of Mycobacterium tuberculosis Controls the SigD Regulon

Journal: Journal of Bacteriology

doi: 10.1128/JB.01537-14

Activation of SigD-dependent transcription requires Rip1 cleavage of RsdA. Quantitative real-time PCR was used to measure the mRNA levels of rpfC (A) or rv1815c (B) in the strains as indicated. Strains were grown in 7H9 medium to log phase for RNA collection.
Figure Legend Snippet: Activation of SigD-dependent transcription requires Rip1 cleavage of RsdA. Quantitative real-time PCR was used to measure the mRNA levels of rpfC (A) or rv1815c (B) in the strains as indicated. Strains were grown in 7H9 medium to log phase for RNA collection.

Techniques Used: Activation Assay, Real-time Polymerase Chain Reaction

25) Product Images from "The Chromosomal parDE2 Toxin–Antitoxin System of Mycobacterium tuberculosis H37Rv: Genetic and Functional Characterization"

Article Title: The Chromosomal parDE2 Toxin–Antitoxin System of Mycobacterium tuberculosis H37Rv: Genetic and Functional Characterization

Journal: Frontiers in Microbiology

doi: 10.3389/fmicb.2016.00886

MParDE2 Promoter identification and regulation. (A) M. smegmatis recombinants viz., MS1, harboring a promoter-less lacZ construct; MS2, containing P 363parD2 -lacZ fusion construct and MS12, vector control strains were grown in 7H9 medium at 37°C and β-galactosidase activity was measured spectrophotometrically, after regular intervals. (B) The transcriptional start site (+1), putative -35 and -10 promoter elements along with parD2 and parE2 translational start sites are indicated in rectangular boxes determined by 5′-RACE PCR analysis. MParD2 amino acid residues substituted with alanine are underlined. (C) Regulation of P parDE2 in M. smegmatis strains. β-galactosidase activity from different M. smegmatis strains harboring 363 bp parDE2 promoter sequence fused with lacZ gene, co-expressing MParD2 or MParDE2 protein complex under the control of groEL promoter. Empty pMV261 vector, parD2 and ParDE2 sequences without promoter were used as negative controls. Error bars indicate mean ± SD from three independent experiments.
Figure Legend Snippet: MParDE2 Promoter identification and regulation. (A) M. smegmatis recombinants viz., MS1, harboring a promoter-less lacZ construct; MS2, containing P 363parD2 -lacZ fusion construct and MS12, vector control strains were grown in 7H9 medium at 37°C and β-galactosidase activity was measured spectrophotometrically, after regular intervals. (B) The transcriptional start site (+1), putative -35 and -10 promoter elements along with parD2 and parE2 translational start sites are indicated in rectangular boxes determined by 5′-RACE PCR analysis. MParD2 amino acid residues substituted with alanine are underlined. (C) Regulation of P parDE2 in M. smegmatis strains. β-galactosidase activity from different M. smegmatis strains harboring 363 bp parDE2 promoter sequence fused with lacZ gene, co-expressing MParD2 or MParDE2 protein complex under the control of groEL promoter. Empty pMV261 vector, parD2 and ParDE2 sequences without promoter were used as negative controls. Error bars indicate mean ± SD from three independent experiments.

Techniques Used: Construct, Plasmid Preparation, Activity Assay, Polymerase Chain Reaction, Sequencing, Expressing

26) Product Images from "Interaction of CarD with RNA Polymerase Mediates Mycobacterium tuberculosis Viability, Rifampin Resistance, and Pathogenesis"

Article Title: Interaction of CarD with RNA Polymerase Mediates Mycobacterium tuberculosis Viability, Rifampin Resistance, and Pathogenesis

Journal: Journal of Bacteriology

doi: 10.1128/JB.00879-12

Weakening the interaction between CarD and the RNAP compromises the survival of mycobacteria during oxidative stress. (A and B) Survival of M. smegmatis strains during oxidative stress. The log-phase M. smegmatis Δ carD attB ∷ tet-carD strains expressing CarD WT , CarD R25E , or CarD R47E in LB were treated for 1 h with either 10 mM or 25 mM H 2 O 2 . After treatment, dilutions were plated on LB. Panel A shows one such experiment, and B graphically represents survival as a ratio of CFU in treated cultures to that in untreated cultures. (C) Survival of the M. tuberculosis strains during oxidative stress. The log-phase M. tuberculosis Δ carD attB ∷ tet-carD strains expressing CarD WT or CarD R47E growing in 7H9 broth were treated for 75 h with 25 mM H 2 O 2 . After treatment, dilutions were plated on 7H10 agar, and survival is graphically represented as the ratio of CFU in treated cultures to that in untreated cultures. The graphs in panels B and C show the mean± standard error of the mean (SEM), and each sample is represented by a black circle. The significance levels in panels B and C were determined by calculating P values by Student's t test; an asterisk indicates significance with a P value of
Figure Legend Snippet: Weakening the interaction between CarD and the RNAP compromises the survival of mycobacteria during oxidative stress. (A and B) Survival of M. smegmatis strains during oxidative stress. The log-phase M. smegmatis Δ carD attB ∷ tet-carD strains expressing CarD WT , CarD R25E , or CarD R47E in LB were treated for 1 h with either 10 mM or 25 mM H 2 O 2 . After treatment, dilutions were plated on LB. Panel A shows one such experiment, and B graphically represents survival as a ratio of CFU in treated cultures to that in untreated cultures. (C) Survival of the M. tuberculosis strains during oxidative stress. The log-phase M. tuberculosis Δ carD attB ∷ tet-carD strains expressing CarD WT or CarD R47E growing in 7H9 broth were treated for 75 h with 25 mM H 2 O 2 . After treatment, dilutions were plated on 7H10 agar, and survival is graphically represented as the ratio of CFU in treated cultures to that in untreated cultures. The graphs in panels B and C show the mean± standard error of the mean (SEM), and each sample is represented by a black circle. The significance levels in panels B and C were determined by calculating P values by Student's t test; an asterisk indicates significance with a P value of

Techniques Used: Expressing

27) Product Images from "Application of Fluorescent Protein Expressing Strains to Evaluation of Anti-Tuberculosis Therapeutic Efficacy In Vitro and In Vivo"

Article Title: Application of Fluorescent Protein Expressing Strains to Evaluation of Anti-Tuberculosis Therapeutic Efficacy In Vitro and In Vivo

Journal: PLoS ONE

doi: 10.1371/journal.pone.0149972

Evaluation of plasmid stability and the absence of effects on bacterial growth. A. Growth curves for M . bovis BCG strains carrying L5-tdTomato, Hsp60-tdTomato or the vector plasmids alone (no fluorescent protein expressed). B. Optical density (OD) of cultures at 600 nm in 7H9 media with (kan 25) or without (no kan) 25 μg/ml kanamycin over 18 days of culture. C. Fluorescence changes of the strain in media with or without kanamycin over 18 days of culture. D. Correlation between fluorescence and OD at 600 nm for the strain grown in medium without kanamycin. E. Correlation between fluorescence and OD at 600 nm for the strain grown in medium with kanamycin. For correlation analyses, samples for OD 600 within the range of 0.05–1 were selected. Data represent one of at least three independent replicate experiments. Values in the Y-axis represent mean of fluorescence or OD values. The error bars are standard deviations.
Figure Legend Snippet: Evaluation of plasmid stability and the absence of effects on bacterial growth. A. Growth curves for M . bovis BCG strains carrying L5-tdTomato, Hsp60-tdTomato or the vector plasmids alone (no fluorescent protein expressed). B. Optical density (OD) of cultures at 600 nm in 7H9 media with (kan 25) or without (no kan) 25 μg/ml kanamycin over 18 days of culture. C. Fluorescence changes of the strain in media with or without kanamycin over 18 days of culture. D. Correlation between fluorescence and OD at 600 nm for the strain grown in medium without kanamycin. E. Correlation between fluorescence and OD at 600 nm for the strain grown in medium with kanamycin. For correlation analyses, samples for OD 600 within the range of 0.05–1 were selected. Data represent one of at least three independent replicate experiments. Values in the Y-axis represent mean of fluorescence or OD values. The error bars are standard deviations.

Techniques Used: Plasmid Preparation, Fluorescence

28) Product Images from "Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿"

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

Journal: Journal of Clinical Microbiology

doi: 10.1128/JCM.01951-10

Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
Figure Legend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

Techniques Used:

Related Articles

High Performance Liquid Chromatography:

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: .. A number of papers examining HPLC methods for the determination of RIF concentrations have been published ( , ), but none have evaluated the stability of RIF in L-J medium and 7H9 broth. .. In our assay, a rapid, accurate, and reproducible HPLC method was developed and validated to measure rifampin concentrations in L-J medium and 7H9 broth medium.

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: .. The calibration curves of RIF in different media showed good linearity over the concentration range of 5 to 50 mg/liter ( r was higher than 0.999 in all cases) ( ).When rifampin was detected by HPLC, the LOD values were 0.08, 0.11, and 0.15 mg/liter and the LOQ values were 0.23, 0.33, and 0.45 mg/liter for L-J medium, 7H9 broth, and RIF stock solution, respectively. .. The analysis showed that coagulation at 85°C for 50 min during the preparation of L-J medium led to about a 28% RIF degradation.

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: .. To answer all these questions, we conducted experiments to serially monitor rifampin stability in L-J medium and in 7H9 broth by high-performance liquid chromatography. .. A number of papers examining HPLC methods for the determination of RIF concentrations have been published ( , ), but none have evaluated the stability of RIF in L-J medium and 7H9 broth.

Clone Assay:

Article Title: Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis
Article Snippet: Bacterial strains and culture conditions M . tuberculosis H37Rv and Mycobacterium smegmatis mc2 155 were cultured in 7H9 broth (Difco) with 0.05% Tween-80 supplemented with 0.2% glycerol and ADC (Bovine Albumin fraction V (50 g/L), dextrose (20 g/L) and catalase (0.0375 g/L)) or Middlebrook OADC (oleic acid ADC), or on Middlebrook 7H10 (Difco) supplemented with ADC or OADC. .. E . coli XL1 Blue strain was used for cloning, while recombinant protein expression was performed using the E . coli Arctic express (DE3) (Agilent Technologies) and the BL21(DE3)pLysS (Novagen) strains.

Selection:

Article Title: Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis
Article Snippet: Bacterial strains and culture conditions M . tuberculosis H37Rv and Mycobacterium smegmatis mc2 155 were cultured in 7H9 broth (Difco) with 0.05% Tween-80 supplemented with 0.2% glycerol and ADC (Bovine Albumin fraction V (50 g/L), dextrose (20 g/L) and catalase (0.0375 g/L)) or Middlebrook OADC (oleic acid ADC), or on Middlebrook 7H10 (Difco) supplemented with ADC or OADC. .. Kanamycin (50 μg/ml), hygromycin (50 μg/ml), 5-Bromo-4 Chloro-3 Indolyl β-D-galactosidase (X-gal) (40 μg/ml) and sucrose (5%) were used for selection purposes when applicable.

Homogenization:

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: .. Calibration curves for the stock solution and 7H9 broth were constructed in a fashion similar to those used with L-J medium but without homogenization/deproteinization. .. Concentrations were derived from a linear regression analysis of the peak area ratio (analyte/IS)-versus-concentration curves.

Mutagenesis:

Article Title: Lansoprazole is an antituberculous prodrug targeting cytochrome bc1
Article Snippet: .. Recombineering method for target confirmation Mtb H37Rv carrying plasmid pJV53 was grown to mid-log phase in 7H9 broth containing 25 μg ml−1 kanamycin and exposed to 0.2% acetamide for 16 h. Competent cells were co-transformed with 100 ng of single-stranded oligos (lagging and leading strand) (5′-CTCACTGCCTGACGACCTGCTGTCGGGACTCGGTCCGCGCGCGGCACTCTCGTCGATCACGCTGGGTATGC-3′—for the L176P mutation) or (5′-GGCGGGCTCGCAGCCAGACTTCTACATGATGTGGGCCGAGGGTCTGGCCCGGATCTGGCCGCCGTGGGAG-3′—for the T313A mutation) and pYUB412 (50 ng) followed by plating on 7H10 agar plates containing 50 μg ml−1 hygromycin. .. SNPs in hygromycin-resistant clones were confirmed by PCR and sequencing using oligos 5′-TGCTGATCACCGGCGTGTAT-3′ and 5′-AAGATGATCCCCGGCAACAG-3′ for L176P or 5′-TTCAAGTCCGGCGCATTTTT-3′ and 5′-TAGACGAACGGCGGCAGAAT-3′ for T313A.

Cell Culture:

Article Title: Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis
Article Snippet: .. Bacterial strains and culture conditions M . tuberculosis H37Rv and Mycobacterium smegmatis mc2 155 were cultured in 7H9 broth (Difco) with 0.05% Tween-80 supplemented with 0.2% glycerol and ADC (Bovine Albumin fraction V (50 g/L), dextrose (20 g/L) and catalase (0.0375 g/L)) or Middlebrook OADC (oleic acid ADC), or on Middlebrook 7H10 (Difco) supplemented with ADC or OADC. .. Kanamycin (50 μg/ml), hygromycin (50 μg/ml), 5-Bromo-4 Chloro-3 Indolyl β-D-galactosidase (X-gal) (40 μg/ml) and sucrose (5%) were used for selection purposes when applicable.

Plasmid Preparation:

Article Title: Lansoprazole is an antituberculous prodrug targeting cytochrome bc1
Article Snippet: .. Recombineering method for target confirmation Mtb H37Rv carrying plasmid pJV53 was grown to mid-log phase in 7H9 broth containing 25 μg ml−1 kanamycin and exposed to 0.2% acetamide for 16 h. Competent cells were co-transformed with 100 ng of single-stranded oligos (lagging and leading strand) (5′-CTCACTGCCTGACGACCTGCTGTCGGGACTCGGTCCGCGCGCGGCACTCTCGTCGATCACGCTGGGTATGC-3′—for the L176P mutation) or (5′-GGCGGGCTCGCAGCCAGACTTCTACATGATGTGGGCCGAGGGTCTGGCCCGGATCTGGCCGCCGTGGGAG-3′—for the T313A mutation) and pYUB412 (50 ng) followed by plating on 7H10 agar plates containing 50 μg ml−1 hygromycin. .. SNPs in hygromycin-resistant clones were confirmed by PCR and sequencing using oligos 5′-TGCTGATCACCGGCGTGTAT-3′ and 5′-AAGATGATCCCCGGCAACAG-3′ for L176P or 5′-TTCAAGTCCGGCGCATTTTT-3′ and 5′-TAGACGAACGGCGGCAGAAT-3′ for T313A.

Concentration Assay:

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: 7H9 broth (Difco) was prepared according to the manufacturer's instruction. .. For the RIF (Sigma, St. Louis, MO)-containing broth preparation, a freshly made RIF– N , N -dimethylformamide (DMF) solution was added to achieve a final concentration of 40 mg/liter.

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: .. The calibration curves of RIF in different media showed good linearity over the concentration range of 5 to 50 mg/liter ( r was higher than 0.999 in all cases) ( ).When rifampin was detected by HPLC, the LOD values were 0.08, 0.11, and 0.15 mg/liter and the LOQ values were 0.23, 0.33, and 0.45 mg/liter for L-J medium, 7H9 broth, and RIF stock solution, respectively. .. The analysis showed that coagulation at 85°C for 50 min during the preparation of L-J medium led to about a 28% RIF degradation.

Incubation:

Article Title: Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp
Article Snippet: All M. tuberculosis strains were derived from Erdman and were grown planktonically at 37°C in 7H9 (broth) or 7H10 (agar) (Difco) medium supplemented with 10% oleic acid-albumin-dextrose-catalase (OADC), 0.5% glycerol, and 0.05% Tween 80 (broth). .. The 24-well dish was placed in a tightly sealed Tupperware dish for 3 weeks, at which point the Tupperware was opened and incubated for another week before photographing.

Article Title: Lansoprazole is an antituberculous prodrug targeting cytochrome bc1
Article Snippet: Culture conditions and REMA assay of other microorganisms Mycobacterium strains were grown in 7H9 broth (Difco) supplemented with Middlebrook ADC enrichment, 0.2% glycerol, 0.05% Tween-80. .. Twofold serial dilutions of each test compound were prepared in 96-well plates containing bacteria in a total volume of 100 μl and then incubated at 37 or 30 °C (as required) before addition of 10 μl of 0.025% resazurin.

other:

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: In summary, our assay provided an analytical method for the determination of rifampin concentrations in L-J medium and 7H9 broth, which may prove useful for monitoring the quality of drug-containing media.

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: Rifampin was not stable in either L-J medium or 7H9 broth.

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: RIF in 7H9 broth and L-J medium was nearly 50% degraded after 1 week of storage at 37°C, and no RIF curve could be detected for L-J medium after 3 weeks of storage at 37°C and for 7H9 brother after 6 weeks.

Construct:

Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿
Article Snippet: .. Calibration curves for the stock solution and 7H9 broth were constructed in a fashion similar to those used with L-J medium but without homogenization/deproteinization. .. Concentrations were derived from a linear regression analysis of the peak area ratio (analyte/IS)-versus-concentration curves.

Expressing:

Article Title: Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis
Article Snippet: Bacterial strains and culture conditions M . tuberculosis H37Rv and Mycobacterium smegmatis mc2 155 were cultured in 7H9 broth (Difco) with 0.05% Tween-80 supplemented with 0.2% glycerol and ADC (Bovine Albumin fraction V (50 g/L), dextrose (20 g/L) and catalase (0.0375 g/L)) or Middlebrook OADC (oleic acid ADC), or on Middlebrook 7H10 (Difco) supplemented with ADC or OADC. .. E . coli XL1 Blue strain was used for cloning, while recombinant protein expression was performed using the E . coli Arctic express (DE3) (Agilent Technologies) and the BL21(DE3)pLysS (Novagen) strains.

Recombinant:

Article Title: Rv1460, a SufR homologue, is a repressor of the suf operon in Mycobacterium tuberculosis
Article Snippet: Bacterial strains and culture conditions M . tuberculosis H37Rv and Mycobacterium smegmatis mc2 155 were cultured in 7H9 broth (Difco) with 0.05% Tween-80 supplemented with 0.2% glycerol and ADC (Bovine Albumin fraction V (50 g/L), dextrose (20 g/L) and catalase (0.0375 g/L)) or Middlebrook OADC (oleic acid ADC), or on Middlebrook 7H10 (Difco) supplemented with ADC or OADC. .. E . coli XL1 Blue strain was used for cloning, while recombinant protein expression was performed using the E . coli Arctic express (DE3) (Agilent Technologies) and the BL21(DE3)pLysS (Novagen) strains.

Derivative Assay:

Article Title: Essential Roles for Mycobacterium tuberculosis Rel beyond the Production of (p)ppGpp
Article Snippet: .. All M. tuberculosis strains were derived from Erdman and were grown planktonically at 37°C in 7H9 (broth) or 7H10 (agar) (Difco) medium supplemented with 10% oleic acid-albumin-dextrose-catalase (OADC), 0.5% glycerol, and 0.05% Tween 80 (broth). ..

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    Difco 7h9 broth
    Serial monitoring outcomes of RIF stability in L-J medium and <t>7H9</t> broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.
    7h9 Broth, supplied by Difco, used in various techniques. Bioz Stars score: 95/100, based on 44 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

    Journal: Journal of Clinical Microbiology

    Article Title: Rifampin Stability in 7H9 Broth and L?wenstein-Jensen Medium ▿

    doi: 10.1128/JCM.01951-10

    Figure Lengend Snippet: Serial monitoring outcomes of RIF stability in L-J medium and 7H9 broth when kept at 37°C and 4°C. (a) Stability of RIF in 7H9 and L-J media when kept at 37°C. (b) Stability of RIF in 7H9 and L-J media when kept at 4°C.

    Article Snippet: Rifampin was not stable in either L-J medium or 7H9 broth.

    Techniques:

    Sources of RNA samples. ( A ) MTB H37Rv was grown in Middlebrook 7H9 medium. Samples were withdrawn for OD 600 measurements and RNA extraction; sigA mRNA and 16S rRNA were quantified by QRT-PCR. •, OD 600 ; □, sigA /16S ratio × 10,000 ± SD ( n ≥ 3). ( B ) C57BL/6 mice were aerosol-infected with ≈500 cfu of MTB H37Rv. Lungs were collected for cfu enumeration and RNA extraction; sigA mRNA and 16S rRNA were quantified by QRT-PCR. •, cfu ± SD ( n ≥ 4); □, sigA /16S ratio × 10,000 ± SD ( n ≥ 3). ( C ) Multilayered organization of granulomatous tissue bordering a 4-cm-diameter cavity in the left upper lung lobe of patient 2 (hematoxylin/eosin-stained). At the periphery were residual airspace and lymphocytic aggregates ( i ). A capsule of fibroblasts and fibrin with scattered epithelioid macrophages ( ii ) surrounded a cellular zone containing clusters of multinucleated giant cells ( iii ). Giant cells were also prominent at the border between the cellular zone and the underlying zone of caseation necrosis ( iv ). The surface of the cavity wall bordering on the necrotic zone showed extensive leukocytic infiltration and numerous acid-fast bacilli, not revealed by hematoxylin/eosin staining ( v ). Magnification: ×4( Left ), ×40 ( Right ). Similar pathology was observed in lung specimens from all four patients analyzed in this study.

    Journal: Proceedings of the National Academy of Sciences of the United States of America

    Article Title: Differential expression of iron-, carbon-, and oxygen-responsive mycobacterial genes in the lungs of chronically infected mice and tuberculosis patients

    doi: 10.1073/pnas.2436197100

    Figure Lengend Snippet: Sources of RNA samples. ( A ) MTB H37Rv was grown in Middlebrook 7H9 medium. Samples were withdrawn for OD 600 measurements and RNA extraction; sigA mRNA and 16S rRNA were quantified by QRT-PCR. •, OD 600 ; □, sigA /16S ratio × 10,000 ± SD ( n ≥ 3). ( B ) C57BL/6 mice were aerosol-infected with ≈500 cfu of MTB H37Rv. Lungs were collected for cfu enumeration and RNA extraction; sigA mRNA and 16S rRNA were quantified by QRT-PCR. •, cfu ± SD ( n ≥ 4); □, sigA /16S ratio × 10,000 ± SD ( n ≥ 3). ( C ) Multilayered organization of granulomatous tissue bordering a 4-cm-diameter cavity in the left upper lung lobe of patient 2 (hematoxylin/eosin-stained). At the periphery were residual airspace and lymphocytic aggregates ( i ). A capsule of fibroblasts and fibrin with scattered epithelioid macrophages ( ii ) surrounded a cellular zone containing clusters of multinucleated giant cells ( iii ). Giant cells were also prominent at the border between the cellular zone and the underlying zone of caseation necrosis ( iv ). The surface of the cavity wall bordering on the necrotic zone showed extensive leukocytic infiltration and numerous acid-fast bacilli, not revealed by hematoxylin/eosin staining ( v ). Magnification: ×4( Left ), ×40 ( Right ). Similar pathology was observed in lung specimens from all four patients analyzed in this study.

    Article Snippet: MTB H37Rv and clinical strains were grown in plastic roller bottles at 37°C in Middlebrook 7H9 broth (Difco) containing 10% oleic acid-albumin-dextrose-catalase (OADC) (Difco), 0.5% glycerol, and 0.05% Tween 80.

    Techniques: RNA Extraction, Quantitative RT-PCR, Mouse Assay, Infection, Staining

    (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook 7H9 medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at

    Journal: Journal of Bacteriology

    Article Title: Organic Hydroperoxide Resistance Protein and Ergothioneine Compensate for Loss of Mycothiol in Mycobacterium smegmatis Mutants ▿ Mutants ▿ †

    doi: 10.1128/JB.01402-10

    Figure Lengend Snippet: (A) Resistance of M. smegmatis to CuOOH is dependent in part upon Ohr. Cells were cultured to late stationary phase (17 days) in Middlebrook 7H9 medium. Cells were then suspended in the same medium at an A 600 of 0.1 and incubated with CuOOH for 2 h at

    Article Snippet: M. smegmatis mc2 155 was grown in Middlebrook 7H9 broth (Difco) with 0.05% Tween 80 and supplemented with either OADC (oleic acid, albumin, glucose, and catalase supplement) or 1% glucose.

    Techniques: Cell Culture, Incubation