Ampicillin

Manufactured by HiMedia

Sourced in India, United States

Ampicillin is a broad-spectrum antibiotic that belongs to the penicillin class. It is a white or almost white crystalline powder that is soluble in water and slightly soluble in alcohol. Ampicillin is commonly used in laboratory settings for various research and experimental purposes.

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Lab products found in correlation

Ciprofloxacin, by HiMedia (57 mentions) Gentamicin, by HiMedia (45 mentions) Chloramphenicol, by HiMedia (41 mentions) Ceftriaxone, by HiMedia (40 mentions) Amikacin, by HiMedia (36 mentions) Tetracycline, by HiMedia (32 mentions) Cefotaxime, by HiMedia (32 mentions) Cotrimoxazole, by HiMedia (31 mentions) Ceftazidime, by HiMedia (30 mentions) Erythromycin, by HiMedia (24 mentions) Imipenem, by HiMedia (24 mentions) Meropenem, by HiMedia (23 mentions) Streptomycin, by HiMedia (21 mentions) Levofloxacin, by HiMedia (20 mentions) Vancomycin, by HiMedia (20 mentions) Ofloxacin, by HiMedia (17 mentions) Gentamycin, by HiMedia (17 mentions) Nalidixic acid, by HiMedia (17 mentions) Piperacillin tazobactam, by HiMedia (15 mentions) Kanamycin, by HiMedia (14 mentions)

Spelling variants of this product

Ampicillin/sulbactam (6 citations) Ampicillin discs (2 citations) Starch ampicillin agar plate (2 citations)

120 protocols using ampicillin

Identification and Phylogenetic Analysis of Aeromonas Isolates

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Isolates were preliminarily screened for Aeromonas by growing in alkaline peptone water enriched with ampicillin (10 μg/mL) (Hi-Media, Maharashtra, India), ampicillin dextrin agar supplemented with ampicillin (10 μg/mL), and vancomycin hydrochloride (2 μg/mL) [15 (link)].
Identification of the strains was confirmed by twenty-five biochemical tests viz., ONPG (β-galactosidase), urease, lysine utilization, nitrate reduction, ornithine utilization, malonate utilization, Voges Proskauer’s (VP), esculin hydrolysis, phenylalanine deamination, citrate utilization, H₂S production, indole, methyl red, oxidase, and the utilization of arabinose, adonitol, xylose, rhamnose, melibiose, cellobiose, saccharose, raffinose, trehalose, glucose, and lactose [35 (link)].
Genomic DNA of the bacterial isolates was extracted using the QIAamp DNA mini kit (Qiagen, Valencia, CA, USA). Aeromonas genus-specific 16S rRNA gene amplification and sequencing was done using a standard protocol [36 (link)]. A combined phylogenetic tree was constructed through the neighbour-joining method using the MEGA X program (Molecular Evolutionary Genetics Analysis, State College, PA, USA) [37 (link)]. Percentage of replicates was shown in the related taxa grouped in the bootstrap analysis (1000 replicates) next to the branches [38 (link)].

Chakraborty N., Das B.K., Bera A.K., Borah S., Mohanty D., Yadav A.K., Kumar J., Koushlesh S.K., Chanu T.N., Panda S.P, & Vallangi R. (2022). Co-Prevalence of Virulence and Pathogenic Potential in Multiple Antibiotic Resistant Aeromonas spp. from Diseased Fishes with In Silico Insight on the Virulent Protein Network. Life, 12(12), 1979.

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Gfp Tagging of Native B. thuringiensis Strain

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Native B. thuringiensis strain VKK-BB2 (GenBank accession number KT714045) was retrieved from the bacterial stock of Insect Physiology and Molecular Biology Laboratory, Division of Entomology, IARI, New Delhi which was originally isolated from the cabbage aphid, Brevicoryne brassicae (Mandla et al. 2017) . The BtVKK-BB2 strain was found to be the most potential out of 12 native strains screened against neonate of BSFB (data not shown here). Thus, BtVKK-BB2 strain was selected for gfp tagging through electroporation. This strain is resistant to ampicillin and penicillin therefore maintained on Luria Bertani agar (LA) supplemented with ampicillin and penicillin (HiMedia Laboratories Pvt Ltd, India) at 50 µg/ml at 30 °C. The Escherichia coli harboring pCAMBIA1302 plasmid (Fig. 1) was grown in Luria Bertani broth (LB) supplemented with 50 µg/ml kanamycin (HiMedia Laboratories Pvt Ltd, India) at 37 °C.

Pola S., Kesharwani A.K., Singh J., Singh D., & Kalia V. (2022). Endophytic ability of indigenous Bacillus thuringiensis strain VKK-BB2: new horizons for the development of novel insect pest-resistant crops. Egyptian Journal of Biological Pest Control, 32(1).

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Characterizing Inducible Gene Expression

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For experimental measurement, the strain was inoculated in LB media (HiMedia) supplemented with Ampicillin (HiMedia, 100μg/ml ) for 16 h. The culture was subsequently diluted 1:200 in fresh M9 minimal media (supplemented with 0.2% casamino acid, 0.4% glucose, 100mM thiamine, 1MMgSO4 , 1MCaCl2 ) and LB media supplemented with Ampicillin (100μg/ml ). The culture was induced with aTc (Sigma‐Aldrich) of concentrations (0, 1, 2 and 4 ng/ml) in different samples and incubated for two hours. Next, different levels (0, 0.2 and 0.4%) of indicated arabinose (HiMedia) were added to the incubated culture. This final culture is placed in three wells of a 96 well clear bottom plate (Eppendorf) and placed in a plate reader (Biotek Synergy H1 multimode) for measurement of optical density (600 nm) and fluorescence (excitation: 485 nm, emission: 525 nm and gain: 60). Based on the plate reader optical density measurement, these cells are in log phase (Supplementary Figure S10). The measurement was taken for ten hours for arabinose co‐variations and twelve hours for aTc co‐variations at 37°C with 5 min sampling and 2 min double orbital shaking between successive readings. The above protocol was repeated for three days.

Patel A, & Sen S. (2020). Experimental evidence for constraints in amplitude‐timescale co‐variation of a biomolecular pulse generating circuit design. IET Systems Biology, 14(5), 217-222.

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Antibiotic Susceptibility Testing of Shigella

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Antibiotic susceptibility testing was done using Kirby-Bauer disc diffusion method (3 ) for the antibiotics: ampicillin 10 µg, trimethoprim-sulphamethoxazole 1.25/23.75 µg, ciprofloxacin 5 µg, ceftriaxone 30 µg, tetracycline 30 µg, and chloramphenicol 30 µg (Himedia Laboratories, Mumbai). The minimum inhibitory concentration (MIC) for ciprofloxacin and ceftriaxone were performed using Epsilometer test (E-test) strips according to the manufacturer's instructions (AB Biomeriuex, India). The inoculum for the susceptibility testing and the interpretation were done as per CLSI (Clinical Laboratory Standards Institute) guidelines (3 ). ATCC Escherichia coli 25922 was used as the control for interpretation of zone diameters. Combination disc method according to CLSI guidelines was used in order to detect ESBL production in ceftriaxone-resistant Shigella isolates.

Sangeetha A.V., Parija S.C., Mandal J, & Krishnamurthy S. (2014). Clinical and Microbiological Profiles of Shigellosis in Children. Journal of Health, Population, and Nutrition, 32(4), 580-586.

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Antimicrobial Resistance Profiling of Virulent Isolates

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The isolates positive for at least one virulence marker gene by PCR were subjected to antimicrobial susceptibility test against the selected antimicrobials (ampicillin-10 μg, amikacin-10 μg, chloramphenicol-30 μg, ceftriaxone-10 μg, cephalexin-30 μg, cipr ofloxacin-10 μg, co-trimoxazole-25 μg, cefoperazone-tazobactam-75 + 10 μg, meropenem-10 μg, norfloxacin-10 μg, gentamicin-10 μg, cefixime-5 μg, doxycycline hydrochloride-10 μg and ofloxacin-5 μg) (HiMedia, India) by disc diffusion method in Mueller–Hinton agar [18 (link)]. The performance of this test was checked by employing E. coli ATCC 25922 as a standard quality control strain. The results were expressed as sensitive, intermediate and resistant as per standard CLSI guidelines [19 ]. MDR was defined as “acquired non-susceptibility to at least one agent in three or more antimicrobial categories” [20 (link)].

Chellapandi K., Dutta T.K., Sharma I., De Mandal S., Kumar N.S, & Ralte L. (2017). Prevalence of multi drug resistant enteropathogenic and enteroinvasive Escherichia coli isolated from children with and without diarrhea in Northeast Indian population. Annals of Clinical Microbiology and Antimicrobials, 16, 49.

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Antibiotic Sensitivity Patterns of Isolates

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In vitro antibiotic sensitivity patterns of the isolates were conducted as per the method of Bauer et al., (1966) . Antibiotics disc (Hi Media Ltd., Mumbai, India) used in the present study were Amikacin (30 mcg), Amoxyclav (30 mcg), Ampicillin (10 mcg), Ciprofloxacin (5 mcg), Colistin (10 mcg), Ceftriaxone (30 mcg), Erythromycin (15 mcg), Enrofloxacin (10 mcg), Gentamicin (10mcg), Neomycin (30 mcg), Penicillin-G (10IU), Streptomycin (10 mcg), Sulphadiazine (300 mcg) and Tetracycline (30 mcg). Diameters of the clear zone of inhibition were measured and the interpretation of the results was made in accordance with the instructions supplied by the manufacturer (Hi Media Ltd., Mumbai, India). Multiple Antibiotic resistance index (MARI) were also determined for each isolates by dividing the number of antibiotics to which the isolate is resistant to by the total numbers of antibiotics tested (Adenaike, 2016) .

Vatalia D., Bhanderi B.B., Nimavat V., & Jhala M.K. (2020). Isolation, Molecular Identification and Multidrug Resistance Profiling of Bacteria Causing Clinical Mastitis in Cows. Agricultural Science Digest - A Research Journal.

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Antibiotic Susceptibility Screening of Isolates

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Antibiotic drug susceptibility was determined by spreading overnight grown culture of the isolates on MRS agar plates as a lawn. Standard antibiotic discs (tetracycline, erythromycin, ampicillin, gentamycin, penicillin, chloramphenicol, cefuroxime, cefoperazone, levofloxacin, norfloxacin, Hi-Media, Mumbai) were placed on the surface of the MRS agar medium aseptically. Plates were incubated for 24 h at 37 °C and observed for zones of inhibition.

Panda S.H., Goli J.K., Das S, & Mohanty N. (2017). Production, optimization and probiotic characterization of potential lactic acid bacteria producing siderophores. AIMS Microbiology, 3(1), 88-107.

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Molecular Cloning and SYBR Green Assay

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Isopropyl alcohol (Analytical Grade)

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Commercially available spin column kits (QiaAmp DNA Blood Mini Kit, Qiagen, GmbH).

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Oligonucleotides were synthesized from BioServe Technologies, Hyderabad, India.

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DMSO, BSA, Tween-20 were procured from Sigma Chemical Co., St. Louis, USA.

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Taq DNA polymerase and pTZ57R/T vector (Thermo Fisher Scientific, USA)

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LB-agar plates, LB broth and ampicillin (Hi-Media, Mumbai, India)

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1X Thermo Scientific Maxima SYBR green master mix (Thermo Fisher Scientific, USA)

Neduvat A.C., Murthy P.M., Sundarrajan S, & Padmanabhan S. (2018). Use of coagulation factor XIII (F13) gene as an internal control for normalization of genomic DNA’s for HLA typing. MethodsX, 5, 881-889.

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Antibiotic Susceptibility of Gram-Negative Bacteria

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The susceptibilities of Gram-negative bacteria (GNB) to the antimicrobial agents ampicillin (10 µg) amoxicillin/clavulanate (30 µg), cefuroxime (30 µg), cefotaxime (30 µg), chloramphenicol (30 µg), cotrimoxazole (25 µg), gentamicin (10 µg), amikacin (30 µg), ciprofloxacin (5 µg), tetracycline (30 µg) and levofloxacin (5 µg) (HiMedia Laboratories, India) were determined with Kirby-Bauer disc diffusion method in accordance with the Clinical and Laboratory Standards Institute (CLSI) guidelines [16 ]. Erythromycin (30 µg), oxacillin (30 µg), and vancomycin (30 µg) were included for Gram positive bacteria (GPB). The reference strain E. coli ATCC 25922 and Staphylococcus aureus ATCC 10221 were included as quality controls in the susceptibility assays. Relative to the panel of antibiotics tested for each isolate, and according to the international standard definitions for acquired resistance, multidrug resistant (MDR) phenotype was defined as in vitro non-susceptibility to ≥1 agent in ≥3 antimicrobial categories [17 (link)]: penicillins, cephalosporins, beta-lactamase inhibitor combinations, fluoroquinolones, aminoglycosides, chloramphenicol, folate pathway inhibitors, tetracyclines, macrolides and glycopeptides.

Obeng-Nkrumah N., Labi A.K., Acquah M.E, & Donkor E.S. (2015). Bloodstream infections in patients with malignancies: implications for antibiotic treatment in a Ghanaian tertiary setting. BMC Research Notes, 8, 742.

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Hemolytic Activity and Antibiotic Susceptibility of LAB

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We assessed the hemolytic activities of LAB strains showing antagonistic activity during initial screening by the presence of α/α (a small zone of greenish-brownish discoloration of the medium, indicating reduction of hemoglobin to methemoglobin), β/β (clear, colorless or light yellow zone surrounding the colonies depicting total lysis of red blood cells), and γ/γ (with no change observed in the medium) hemolysis following the protocol of Maragkoudakis et al. (2006) (link). In the antibiotic susceptibility test, we examined promising screened and characterized LAB isolates using the agar disc diffusion method (Bauer et al., 1966) (link). Twelve antibiotics were tested (Hi-Media): penicillin G (2 units), ceftriaxone (30 µg), ampicillin (25 µg), vancomycin (30 µg), oxacillin (1 µg), streptomycin (10 µg), chloramphenicol (30 µg), gentamicin (10 µg), erythromycin (10 µg), tetracycline (10 µg), novobiocin (30 µg), and ciprofloxacin (10 µg). Isolates were categorized as sensitive (≥21 mm), intermediate (16-20 mm), or resistant (≤15 mm) (Liasi et al., 2009) . The multiple antibiotics resistance (MAR) index was determined for each probiotic strain as previously described by Ngwai et al. (2011) .

Reuben R.C., Roy P.C., Sarkar S.L., Rubayet Ul Alam A.S.M, & Jahid I.K. (2020). Characterization and evaluation of lactic acid bacteria from indigenous raw milk for potential probiotic properties. Journal of dairy science, 103(2).

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