Most cell lines were originally purchased from the American Type Culture Collection (ATCC) and were cultured as per the standard ATCC protocols. LNCaP-AR and LAPC4 cells were gifts from Dr. Charles Sawyers lab (Memorial Sloan-Kettering Cancer Center, New York, NY). Until otherwise stated, for all the experiments LNCaP, PNT2, LNCaP-AR, C42B, 22RV1, DU145, PC3 cells were grown in the RPMI 1640 medium (Gibco) and VCaP cells in the DMEM with Glutamax (Gibco) medium supplemented with 10% Full Bovine Serum (FBS; Invitrogen). LAPC4 cells were grown in IMEM (Gibco) medium supplemented with 15%FBS and 1nM of R1881. Immortalized normal prostate cells: RWPE1 were grown in keratinocyte media with regular supplements (Lonza); PNT2 were grown in RPMI medium with 10%FBS. HEK293 cells were grown in DMEM (Gibco) medium with 10% FBS. All cells were grown in a humidified 5%CO2 incubator at 37℃. All cell lines were biweekly tested to be free of mycoplasma contamination and genotyped every month at the University of Michigan Sequencing Core using Profiler Plus (Applied Biosystems) and compared with corresponding short tandem repeat (STR) profiles in the ATCC database to authenticate their identity in culture between passages and experiments.
Profiler plus
Manufactured by Thermo Fisher Scientific
Sourced in United States
The Profiler Plus is a laboratory instrument designed for DNA profiling and analysis. It provides a comprehensive solution for the identification and comparison of DNA samples. The core function of the Profiler Plus is to generate DNA profiles from biological samples, which can be used for various applications, including forensics, paternity testing, and human identification.
Automatically generated - may contain errors
Lab products found in correlation
Dulbecco modified eagle medium (dmem), by Thermo Fisher Scientific (3 mentions)
Glutamax, by Thermo Fisher Scientific (3 mentions)
Regular supplements, by Lonza (2 mentions)
Full bovine serum fbs, by Thermo Fisher Scientific (2 mentions)
Lapc4, by Thermo Fisher Scientific (2 mentions)
Rwpe1, by Lonza (2 mentions)
Rpmi 1640 medium, by Thermo Fisher Scientific (2 mentions)
Rmpi 1640, by Thermo Fisher Scientific (1 mentions)
Fetal bovine serum (fbs), by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
3130xl genetic analyser, by Thermo Fisher Scientific (1 mentions)
6 protocols using profiler plus
1
Prostate Cancer Cell Line Cultivation
2
Cell Line Validation and Maintenance
3
Standard Cell Line Culture Conditions
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Cell lines were maintained using standard conditions. Specifically, PC3 and LNCaP cell lines [American Type Culture Collection (ATCC), Manassas, VA] were cultured in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) (Gibco; Thermo Fisher Scientific). All of the cell lines were grown at 37°C in a 5% CO2 cell culture incubator. To ensure identity, the cell lines were genotyped at the University of Michigan Sequencing Core using Profiler Plus (Applied Biosystems, Foster City, CA) and compared with the short tandem repeat profiles of respective cell lines available in the STR Profile Database (ATCC). All of the cell lines were routinely tested and found to be free of Mycoplasma contamination.
4
Prostate Cancer Cell Line Cultivation
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Most cell lines were originally purchased from the American Type Culture Collection (ATCC) and were cultured as per the standard ATCC protocols. LNCaP-AR and LAPC4 cells were gifts from Dr. Charles Sawyers lab (Memorial Sloan-Kettering Cancer Center, New York, NY). Until otherwise stated, for all the experiments LNCaP, PNT2, LNCaP-AR, C42B, 22RV1, DU145, PC3 cells were grown in the RPMI 1640 medium (Gibco) and VCaP cells in the DMEM with Glutamax (Gibco) medium supplemented with 10% Full Bovine Serum (FBS; Invitrogen). LAPC4 cells were grown in IMEM (Gibco) medium supplemented with 15%FBS and 1nM of R1881. Immortalized normal prostate cells: RWPE1 were grown in keratinocyte media with regular supplements (Lonza); PNT2 were grown in RPMI medium with 10%FBS. HEK293 cells were grown in DMEM (Gibco) medium with 10% FBS. All cells were grown in a humidified 5%CO2 incubator at 37℃. All cell lines were biweekly tested to be free of mycoplasma contamination and genotyped every month at the University of Michigan Sequencing Core using Profiler Plus (Applied Biosystems) and compared with corresponding short tandem repeat (STR) profiles in the ATCC database to authenticate their identity in culture between passages and experiments.
5
Cell Line Authentication and Maintenance
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Most cell lines were originally purchased from the American Type Culture Collection (ATCC) and were cultured as per standard ATCC protocols. Unless otherwise stated, for all experiments, LNCaP and 22RV1 cells were grown in RPMI 1640 medium (Gibco) and VCaP cells in DMEM with Glutamax (Gibco) medium supplemented with 10% full bovine serum (FBS; Invitrogen). HEK293 cells were grown in DMEM (Gibco) medium with 10% FBS. All cells were grown in a humidified 5% CO2 incubator at 37 °C. All cell lines were tested once a fortnight to be free of mycoplasma contamination and genotyped every month at the University of Michigan Sequencing Core using Profiler Plus (Applied Biosystems) and compared with corresponding short tandem repeat profiles in the ATCC database to authenticate their identity in culture between passages and experiments.
6
Microsatellite Genotyping Protocol
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
Amplification was carried out using Profiler Plus (Applied Biosystems, USA). Cycling was performed on a 9700 GeneAmp cycler and consisted of an initial denaturation at 95°C for 10 min followed by 28 cycles of 94°C for 1 min, 59°C for 1 min and 72°C for 1 min, followed by a final extension at 60°C for 45 min. All reactions were initially performed in 25 μL volumes, with 10 μL of DNA (diluted when appropriate to allow a final concentration of ideally between 0.5 and 1 ng). Capillary electrophoresis was performed using an Applied Biosystems 3130xl genetic analyser and profiling analysis was undertaken using GeneMapper ID v3.2.1. The analysis was repeated in duplicate or triplicate, in 12.5 μL reaction volumes including 5 μL of DNA, when peak heights were lower than 50 Relative Fluroscence Units (RFU). A consensus approach was then used to identify peaks from the multiple reactions
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!