N garde mycoplasma pcr reagent

Manufactured by Euroclone

The N-GARDE Mycoplasma PCR Reagent is a molecular diagnostic tool used for the detection of Mycoplasma species in cell cultures. It is designed to amplify specific DNA sequences from Mycoplasma organisms through the polymerase chain reaction (PCR) process.

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Lab products found in correlation

Fetal bovine serum (fbs), by Euroclone (3 mentions) Williams e medium, by Merck Group (2 mentions) Dulbecco s modified eagle s medium, by Thermo Fisher Scientific (2 mentions) Rpmi medium, by Euroclone (2 mentions) L glutamine, by Euroclone (2 mentions) Penicillin streptomycin, by Euroclone (1 mentions) Mirneasy kit, by Qiagen (1 mentions) Dharmafect transfection reagent, by Horizon Discovery (1 mentions) Dulbecco s modified eagle s medium, by Euroclone (1 mentions) Hct116, by American Type Culture Collection (1 mentions) Penicillin, by Thermo Fisher Scientific (1 mentions) Mcf 7 cell line, by Merck Group (1 mentions) Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions) Geneticin, by Thermo Fisher Scientific (1 mentions) Blasticidin, by Thermo Fisher Scientific (1 mentions) Doxycycline, by Merck Group (1 mentions) Zeocine, by Thermo Fisher Scientific (1 mentions) Penicillin, by Euroclone (1 mentions) Alpha mem, by Euroclone (1 mentions)

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N-GARDE Mycoplasma PCR Reagent set (9 citations)

3 protocols using n garde mycoplasma pcr reagent

Inhibition of APE1 Endonuclease and Redox Activities in Cell Lines

Cited 2 times

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A549 and CH12F3 cells were grown in RPMI medium (Euroclone, Milan, Italy), HeLa clones in Dulbecco's modified Eagle's medium (Invitrogen, Monza, Italy) while JHH-6 were cultured in William’s medium E (Sigma-Aldrich, St. Louis, MO). CH12F3 containing two (+ / + /Δ) and zero copies of APE1 (Δ/Δ/Δ) have been described previously [27 (link)]. All cells were supplemented with 10% fetal bovine serum (Euroclone), 1% penicillin–streptomycin solution (100 U/mL penicillin, 100 mg/mL streptomycin), 2 mM L-glutamine (Euroclone) and cultured in a humidified incubator at 5% CO₂ at 37 °C. Cells were tested as free of mycoplasma contamination (N-GARDE Mycoplasma PCR Reagent, Euroclone).
For APE1 endonuclease activity inhibition, A549 cells were treated with 20 µM APE1 endonuclease inhibitor #3 [28 (link)] while 100 µM of E3330 [29 (link)] was used for redox activity inhibition.

Antoniali G., Dalla E., Mangiapane G., Zhao X., Jing X., Cheng Y., De Sanctis V., Ayyildiz D., Piazza S., Li M, & Tell G. (2022). APE1 controls DICER1 expression in NSCLC through miR-33a and miR-130b. Cellular and Molecular Life Sciences, 79(8), 446.

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Silencing of hAPE1 in cancer cells

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JHH-6 hepatocarcinoma cells23 (link) were grown in William’s medium E (Sigma-Aldrich, St. Louis, MO, USA). Huh7 differentiated hepatocytes derived from cellular carcinoma,24 (link) HepG2 differentiated hepatocellular carcinoma cells,25 (link) and HCT-116 cells were grown in Dulbecco’s modified Eagle’s medium (EuroClone, Milan, Italy). A549 cells were grown in RPMI medium (Euroclone, Milan, Italy). All media were supplemented with 10% fetal bovine serum (Euroclone), 2 mM L-glutamine (Euroclone), 100 U/mL penicillin, and 100 mg/mL streptomycin. Cells were negative for mycoplasma. The tests were performed with N-GARDE Mycoplasma PCR Reagent (Euroclone).
Cells were seeded in the number of 1.0×10⁶ in a 10 cm dish. The following day, transfection was performed using 100 pmol of custom hAPE1 siRNA or with nontargeting siRNA pool (siSCR) as a control (GE Healthcare Dharmacon, Lafayette, CO, USA). The DharmaFECT transfection reagent (GE Healthcare Dharmacon) was employed for transfection following the manufacturer’s protocols. Two days after transfection, cells were collected, and whole cell extracts and RNA were prepared. RNA extraction was carried out using miRNeasy kit (Qiagen, Germantown, MD, USA) following the manufacturer’s protocols.

Mangiapane G., Pascut D., Dalla E., Antoniali G., Degrassi M., Crocè L.S., De Sanctis V., Piazza S., Canarutto G., Tiribelli C, & Tell G. (2023). Clinical Significance of Apurinic/Apyrimidinic Endodeoxyribonuclease 1 and MicroRNA Axis in Hepatocellular Carcinoma. Journal of Clinical and Translational Hepatology, 11(6), 1291-1307.

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Cell Culture Protocols for Cancer Research

Cited 1 time

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HeLa, HCT-116 (ATCC, Manassas, VA), and MCF-7 cell lines (Sigma-Aldrich, St. Louis, MO) were grown in Dulbecco's modified Eagle's medium (Invitrogen, Monza, Italy) supplemented with 10% fetal bovine serum (Euroclone, Milan, Italy), 100 U ml⁻¹ penicillin, 10 μg ml⁻¹ streptomycin sulphate. OCI/AML-2 and OCI/AML-3 cell lines (a kind gift by Emanuela Colombo) were grown in alpha-MEM (Euroclone) supplemented with 20% fetal bovine serum, 100 U ml⁻¹ penicillin and 10 μg ml⁻¹ streptomycin sulfate. HeLa cell clones expressing an ectopic APE1–FLAG-tagged form^{13 (link)} were grown in Dulbecco's modified Eagle's medium supplemented with 10% fetal bovine serum, 100 U ml⁻¹ penicillin, 10 μg ml⁻¹ streptomycin sulfate, 3 μg ml⁻¹ blasticidin, 100 μg ml⁻¹ zeocine, and 400 μg ml⁻¹ geneticin (Invitrogen, Carlsbad, CA). For inducible APE1-shRNA experiments, doxycycline (1 μg ml⁻¹; Sigma-Aldrich) was added to the cell culture medium, and cells were grown for 10 days, as previously described^{13 (link), 14 (link)}. All cell lines were tested and free of mycoplasma contamination (N-GARDE Mycoplasma PCR Reagent, Euroclone).

Antoniali G., Serra F., Lirussi L., Tanaka M., D’Ambrosio C., Zhang S., Radovic S., Dalla E., Ciani Y., Scaloni A., Li M., Piazza S, & Tell G. (2017). Mammalian APE1 controls miRNA processing and its interactome is linked to cancer RNA metabolism. Nature Communications, 8, 797.

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