Mycoplasma plus pcr primer set

Manufactured by Agilent Technologies

Sourced in United States

The Mycoplasma Plus PCR Primer Set is a laboratory equipment product designed for the detection of mycoplasma contamination in cell cultures. It provides a reliable and efficient method for the identification of various mycoplasma species.

Automatically generated - may contain errors

Lab products found in correlation

31 protocols using mycoplasma plus pcr primer set

Culturing A549 Cells for Research

Check if the same lab product or an alternative is used in the 5 most similar protocols

The A549 cell line was acquired from the American Type Culture Collective (ATCC) (Manassas, Virginia, USA) and authenticated by Genetic Testing Biotechnology Corporation (Suzhou, China) using short tandem repeat (STR) profiling. Mycoplasma contamination was tested using the Mycoplasma Plus PCR Primer Set (Agilent, Santa Clara, USA) and were found to be negative. Cells were cultured in Dulbecco's Modified Eagle's Medium (Gibco, New York, USA) containing 10% fetal bovine serum (Gibco, New York, USA) and a mixture of 100 units/mL penicillin and 100 μg/mL streptomycin (Gibco) at 37 °C and 5% CO₂.

Zhang X., Li G., Guo Y., Song Y., Chen L., Ruan Q., Wang Y., Sun L., Hu Y., Zhou J., Ren B, & Guo J. (2019). Regulation of ezrin tension by S-nitrosylation mediates non-small cell lung cancer invasion and metastasis. Theranostics, 9(9), 2555-2571.

+ Open protocol

+ Expand

Cell Culture Protocols for Osteosarcoma and Keratinocytes

Check if the same lab product or an alternative is used in the 5 most similar protocols

U2OS osteosarcoma cells (American Type Culture Collection, Manassas, VA) and HaCaT (immortalized skin keratinocytes) were grown in Dulbecco’s minimal essential medium with GlutaMAX (Invitrogen, Carlsbad, CA) and 10 % fetal bovine serum. OKF6-TERT2 cells (TERT-immortalized primary oral keratinocytes) were cultured in keratinocyte serum-free media (Invitrogen) supplemented with growth factors as per the manufacturer’s recommendations. The media were supplemented with 100 IU/mL penicillin and 100 IU/mL streptomycin (Invitrogen). Quality control was sporadically performed using the Mycoplasma Plus™ PCR Primer Set (Agilent Technologies, Santa Clara, CA).

Lin P., Mobasher M.E., Hakakian Y., Kakarla V., Naseem A.F., Ziai H, & Alawi F. (2015). Differential requirements for H/ACA ribonucleoprotein components in cell proliferation and response to DNA damage. Histochemistry and cell biology, 144(6), 543-558.

+ Open protocol

+ Expand

Cell Line Authentication and Maintenance

Check if the same lab product or an alternative is used in the 5 most similar protocols

Raji and THP-1 cell line were provided by Dr. Francisco Hernandez-Ilizaliturri (Roswell Park Cancer Institute). Both cell lines were tested and authenticated for mycoplasma using the Agilent Mycoplasma Plus PCR Primer Set before experimental use. Cells were maintained in media composed of RPMI-1640, 10% FBS and 1% penicillin & streptomycin. Cells were passaged using 75 cm² flasks at 37°C and 5% CO₂.

Mohammadpour H., Tsuji T., MacDonald C.R., Sarow J.L., Rosenheck H., Daneshmandi S., Choi J.E., Qiu J., Matsuzaki J., Witkiewicz A.K., Attwood K., Blazar B.R., Odunsi K., Repasky E.A, & McCarthy P.L. (2023). Galectin-3 expression in donor T cells reduces GvHD severity and lethality after allogeneic hematopoietic cell transplantation. Cell reports, 42(3), 112250.

+ Open protocol

+ Expand

Cell Line Characterization and Validation

Check if the same lab product or an alternative is used in the 5 most similar protocols

293T and PC9 cells were purchased from ATCC (in 2009) and Sigma Aldrich (in 2014), respectively. Ba/F3 cells were a generous gift from the lab of Dr. David Weinstock (in 2014). 293T cells were cultured in DMEM, supplemented with 10% FBS, streptomycin and penicillin. PC9 and transformed Ba/F3 cells were maintained in RPMI1640 supplemented with 10% FBS, streptomycin and penicillin. The human 293T and PC9 cell lines were authenticated using the Promega GenePrint 10 System at the RTSF Genomics Core at Michigan State University in August 2016. Ba/F3 cells were not authenticated, since their STR profile has not been made publicly available. All cell lines used in the study tested negative for mycoplasma as determined by the Mycoplasma Plus PCR Primer Set (Agilent) in August 2016.

Bahcall M., Sim T., Paweletz C.P., Patel J.D., Alden R.S., Kuang Y., Sacher A.G., Kim N.D., Lydon C.A., Awad M.M., Jaklitsch M.T., Sholl L.M., Jänne P.A, & Oxnard G.R. (2016). Acquired MET D1228V mutation and resistance to MET inhibition in lung cancer. Cancer discovery, 6(12), 1334-1341.

+ Open protocol

+ Expand

Lentiviral Knockdown of LTBP4 in Glioma Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

U87 (ATCC HTB-14) cell line was acquired through American Type Culture Collection. U251 (Sigma, catalogue number 09063001) cell line was obtained through Sigma. Cell lines were cultured in DMEM supplemented with 10% fetal bovine serum (FBS, Sigma). Cells were routinely tested for mycoplasma contamination using Mycoplasma Plus PCR Primer Set (Agilent, Santa Clara, CA) and were found to be negative.
Lentivirus was generated by co-transfection of the lentiviral vectors with pCMV-ΔCMV-ors wpMD2.G plasmids into HEK293T cells as previously described (Niola et al. JCI 2013; Carro et al. Nature 2010). shRNA sequences for LTBP4 are in Supplementary Table 11.
After infection cells were selected with Puromycin (Sigma) at concentration of 2 mg/ml for 48 h. Cells were analysed by western blot, qRT-PCR and growth assay 3 days later.
Evaluation of cell growth was performed using the MTT assay. Cells were plated at density of 2.5×10³ cells/well into 96 well plates in 6 replicates and allowed to adhere for 24 h. Viability was assessed daily by adding MTT ((3-[4,5-dimethylthiazol-2-yl]-2,5- diphenyltetrazolium, Sigma 5mg/ml in PBS). Following 4h incubation period, medium was removed and formazan crystal were solubilized with acidic isopropanol (0.1 N HCl in absolute isopropanol. The absorbance at 550 nm was measured with a plate reader.

Wang J., Cazzato E., Ladewig E., Frattini V., Rosenbloom D.I., Zairis S., Abate F., Liu Z., Elliott O., Shin Y.J., Lee J.K., Lee I.H., Park W.Y., Eoli M., Blumberg A.J., Lasorella A., Nam D.H., Finocchiaro G., Iavarone A, & Rabadan R. (2016). Clonal Evolution of Glioblastoma under Therapy. Nature genetics, 48(7), 768-776.

+ Open protocol

+ Expand

Mycoplasma Detection and Identification

Check if the same lab product or an alternative is used in the 5 most similar protocols

The Mycoplasma Plus PCR primer set (Agilent) was used to detect mycoplasma infections in our cell cultures. If the cells are infected with mycoplasma, the primers yield a single 874 bp amplification product, regardless of which species of mycoplasma is present in the sample. Restriction-fragment analysis of the amplification products using the restriction enzyme Sau3A I determined which species of mycoplasma was present in our samples. A species analysis was conducted by digesting a portion of the PCR sample, separating the restriction fragments using agarose gel electrophoresis and comparing the fragmentation pattern to the fingerprints that identify the mycoplasma species.

Boslett B., Nag S, & Resnick A. (2014). Detection and Antibiotic Treatment of Mycoplasma arginini Contamination in a Mouse Epithelial Cell Line Restore Normal Cell Physiology. BioMed Research International, 2014, 532105.

+ Open protocol

+ Expand

Cell Culture and Drug Treatments

Check if the same lab product or an alternative is used in the 5 most similar protocols

SW1271, H1299, H2087, H2347, H69, H82, H209, and Glc16 were purchased from American Type Culture Collection (ATCC). All cell lines except SW1271 were maintained in Roswell Park Memorial Institute (RPMI)‐1640 (Gibco, Thermo Fisher Scientific, Waltham, MA, USA) supplemented with 10% FBS, 100 U·mL⁻¹ penicillin, and 100 µg·mL⁻¹ streptomycin (Gibco, Thermo Fisher Scientific). SW1271 was grown in Dulbecco’s modified Eagle medium (Gibco, Thermo Fisher Scientific) with 10% FBS. Trametinib (Yamaguchi et al., 2011), selumetinib (Davis et al., 2011), INK128 (Hsieh et al., 2012), AZD8055 (Chresta et al., 2010), PI103 (Raynaud et al., 2009), BKM120 (Burger et al., 2011), ZSTK474 (Kong and Yamori, 2007), and MK2206 (Hirai et al., 2010) were purchased from Selleck Chemicals (Houston, TX, USA). Torin2 was synthesized using previously published methods (Liu et al., 2013). Stock solutions of all drugs were prepared at 10 mm in DMSO (Sigma‐Aldrich, St. Louis, MO, USA) and stored at −80 °C. Cell lines were authenticated by single tandem repeat analysis at Michigan State University in October 2017 and tested negative for mycoplasma as determined by the Mycoplasma Plus PCR Primer Set (Agilent Technologies, Santa Clara, CA, USA).

Ogino A., Choi J., Lin M., Wilkens M.K., Calles A., Xu M., Adeni A.E., Chambers E.S., Capelletti M., Butaney M., Gray N.S., Gokhale P.C., Palakurthi S., Kirschmeier P., Oxnard G.R., Sholl L.M, & Jänne P.A. (2020). Genomic and pathological heterogeneity in clinically diagnosed small cell lung cancer in never/light smokers identifies therapeutically targetable alterations. Molecular Oncology, 15(1), 27-42.

+ Open protocol

+ Expand

U2OS Cell Culture Protocol

Check if the same lab product or an alternative is used in the 5 most similar protocols

U2OS cells (American Type Culture Collection, Manassas, VA) were grown in Dulbecco’s minimal essential medium with GlutaMAX (Invitrogen, Carlsbad, CA) and 10% fetal bovine serum with 100 IU/mL penicillin and 100 IU/mL streptomycin (Invitrogen). Quality control was sporadically performed using the Mycoplasma Plus™ PCR Primer Set (Agilent Technologies, Santa Clara, CA).

Lin P., Mobasher M.E, & Alawi F. (2014). ACUTE DYSKERIN DEPLETION TRIGGERS CELLULAR SENESCENCE AND RENDERS OSTEOSARCOMA CELLS RESISTANT TO GENOTOXIC STRESS-INDUCED APOPTOSIS. Biochemical and biophysical research communications, 446(4), 1268-1275.

+ Open protocol

+ Expand

Culturing Human Brain Endothelial Cells

Check if the same lab product or an alternative is used in the 5 most similar protocols

The human brain microvascular endothelial cell (HBMEC) line was acquired from the Bena Culture Collection (BNCC, Suzhou, China) and authenticated by the Genetic Testing Biotechnology Corporation (Suzhou, China) by using short tandem repeats (STR). Mycoplasma Plus PCR Primer Set (Agilent, Santa Clara, USA) was used to test for mycoplasma contamination, and results were negative. HBMEC cells were cultured in Dulbecco’s Modified Eagle’s Medium (Gibco, New York, USA) containing 10% fetal bovine serum (Gibco, New York, USA), 100 units/mL penicillin, and 100 μg/mL streptomycin (Gibco) and maintained in a 5% CO₂ incubator at 37°C.

Li C., Chen L., Wang Y., Wang T., Di D., Zhang H., Zhao H., Shen X, & Guo J. (2021). Protein Nanoparticle-Related Osmotic Pressure Modifies Nonselective Permeability of the Blood–Brain Barrier by Increasing Membrane Fluidity. International Journal of Nanomedicine, 16, 1663-1680.

+ Open protocol

+ Expand

Amplification and Titration of Influenza Viruses

Check if the same lab product or an alternative is used in the 5 most similar protocols

Mouse-adapted X-79 (H3N2) (A/Philippines/2/1982 (H3N2): A/Puerto Rico/8/1934 (H1N1)) virus was amplified in the lungs of 4 week-old BALB/c mice (Taconic).^{43 (link)} A/Anhui/1/2013 (H7N9) was provided by the Centers for Disease Control (Atlanta, GA) and was grown in embryonated hen’s eggs. Virus stocks, nasal washes, and tissue homogenates were titrated on MDCK cells (ATCC), and virus titers were expressed as median tissue culture infectious dose (TCID₅₀). MDCK cells were confirmed to be mycoplasma-free using the Mycoplasma Plus PCR Primer Set (Agilent Technologies).

Sutton T.C., Chakraborty S., Mallajosyula V.V., Lamirande E.W., Ganti K., Bock K.W., Moore I.N., Varadarajan R, & Subbarao K. (2017). Protective efficacy of influenza group 2 hemagglutinin stem-fragment immunogen vaccines. NPJ Vaccines, 2, 35.

+ Open protocol

+ Expand

About PubCompare

Our mission is to provide scientists with the largest repository of trustworthy protocols and intelligent analytical tools, thereby offering them extensive information to design robust protocols aimed at minimizing the risk of failures.

We believe that the most crucial aspect is to grant scientists access to a wide range of reliable sources and new useful tools that surpass human capabilities.

However, we trust in allowing scientists to determine how to construct their own protocols based on this information, as they are the experts in their field.

Ready to get started?

Revolutionizing how scientists
search and build protocols!