The murine Ret melanoma cell line was established from skin melanomas isolated from RET transgenic mice16 (link) and cultured in RPMI-1640 with GlutaMAX (Thermo Fisher) and supplemented with 10% heat-inactivated FBS (Merck) and 1% penicillin/streptomycin (Thermo Fisher). The immortalized myeloid suppressor cell line MSC-221 (link) was provided by Dr. S. Ugel (University of Verona, Italy) and cultured in RPMI-1640 with GlutaMAXTM and supplemented with 10 mM sodium pyruvate (Thermo Fisher), 10% heat-inactivated FBS and 1% penicillin/streptomycin. Cell lines were maintained under 5% CO2 at 37°C and routinely tested for Mycoplasma contamination using the Mycoplasma Detection Kit for Conventional PCR (Minerva Biolabs). Different cytokines, chemokines and growth factors (PeproTech) and TLR ligands (InvivoGen) were used for cell stimulation (online supplementary table S1 ).
Mycoplasma detection kit for conventional pcr
Manufactured by Minerva Biolabs
Sourced in Germany
The Mycoplasma Detection Kit for Conventional PCR is a laboratory tool designed to detect the presence of mycoplasma contamination in cell cultures. The kit utilizes the conventional polymerase chain reaction (PCR) method to amplify and identify specific mycoplasma DNA sequences.
Automatically generated - may contain errors
Lab products found in correlation
Penicillin streptomycin, by Thermo Fisher Scientific (2 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (2 mentions)
Glutamax, by Thermo Fisher Scientific (2 mentions)
Glutamaxtm, by Thermo Fisher Scientific (1 mentions)
Sodium pyruvate, by Thermo Fisher Scientific (1 mentions)
Heat inactivated, by Merck Group (1 mentions)
Tlr ligands, by InvivoGen (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Bact alert, by bioMérieux (1 mentions)
2 protocols using mycoplasma detection kit for conventional pcr
1
Murine Ret Melanoma and Myeloid Suppressor Cell Culture
2
Microbiological Screening of Cell Cultures
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
For microbiological control, the samples (HPL, media and cellular supernatants) were tested through BacT/ALERT (bioMérieux, Durham, NC, USA) (26) (link) by the Bacteriology Laboratory of the City of Science and Health of Turin, S. Anna Hospital.
We performed the mycoplasma detection test (Mycoplasma Detection Kit for conventional PCR, Minerva Biolabs, Berlin, Germany), with the use of an aliquot of supernatant at each passage during the expansion process and on an amount of each growth supplement (pure HPL, iHPL and FBS). Briefly, any mycoplasma contamination of the analyzed samples was detected through a semi-quantitative PCR reaction. As the positive control, we used a positive control DNA, provided by the kit. To monitor the success of the extraction procedure, we used the internal control DNA of Venor GeM. As a negative control, we used PCR-grade water.
We performed the mycoplasma detection test (Mycoplasma Detection Kit for conventional PCR, Minerva Biolabs, Berlin, Germany), with the use of an aliquot of supernatant at each passage during the expansion process and on an amount of each growth supplement (pure HPL, iHPL and FBS). Briefly, any mycoplasma contamination of the analyzed samples was detected through a semi-quantitative PCR reaction. As the positive control, we used a positive control DNA, provided by the kit. To monitor the success of the extraction procedure, we used the internal control DNA of Venor GeM. As a negative control, we used PCR-grade water.
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?
Sign up for free.
Registration takes 20 seconds.
Available from any computer
No download required
Revolutionizing how scientists
search and build protocols!