Frozen vials of the metastatic melanoma cell-line COLO829 (CRL-1974) and EBV transformed B lymphoblast cells from the same individual (COLO829BL, CRL-1980) were purchased from ATCC via Cedarlane (Burlington, Canada). COLO829BL cells were cultured to passage #3 at 37 °C in RPMI-1640 media supplemented with Fetal Bovine Serum (10%). COLO829 cells were cultured to passage #14 at 37 °C in RPMI-1640 media supplemented with Fetal Bovine Serum (10%).
Colo829bl
Manufactured by American Type Culture Collection
Sourced in Canada
The COLO829BL is a laboratory cell line derived from a human melanoma. It is a suspension cell line that can be used for various cell-based assays and experiments.
Automatically generated - may contain errors
Lab products found in correlation
Colo829, by American Type Culture Collection (9 mentions)
Gotaq dna polymerase, by Promega (2 mentions)
Colo 829, by Merck Group (2 mentions)
Genome dna kit, by Thermo Fisher Scientific (2 mentions)
Rpmi 1640, by Merck Group (2 mentions)
Hcc1187, by American Type Culture Collection (1 mentions)
Hcc 1143 bl, by American Type Culture Collection (1 mentions)
Hcc 1187bl, by American Type Culture Collection (1 mentions)
Hcc1143, by American Type Culture Collection (1 mentions)
Mitochondrial isolation kit, by Miltenyi Biotec (1 mentions)
Qproteome kit, by Qiagen (1 mentions)
Dneasy blood and tissue kit, by Qiagen (1 mentions)
Countess cell counter, by Thermo Fisher Scientific (1 mentions)
Rpmi 1640, by Thermo Fisher Scientific (1 mentions)
Antibiotic antimycotic, by Thermo Fisher Scientific (1 mentions)
Dmem media, by Thermo Fisher Scientific (1 mentions)
9 protocols using colo829bl
1
Culturing Metastatic Melanoma and EBV B Cells
2
Karyotyping Cancer Cell Lines
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The cancer cell lines (COLO-829 ATCC CRL-1974, COLO-829BL ATCC CRL-1980, HCC-1143 ATCC CRL-2321, HCC-1143 BL ATCC CRL-2362, HCC-1187 ATCC CRL-2322 and HCC-1187BL ATCC CRL-2323) were obtained from ATCC18 . The cell lines were cultured using the recommendations from ATCC. Cultured cells were split into two aliquots for metaphase chromosome preparation and karyotype analysis. Representative images and karyotypes are reported in Supplemental Fig. S1 . The number of passages is indicated in Supplemental Table 1 .
3
Benchmarking DNA Sequencing Technologies
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COLO829 (ATCC® CRL-1974™) and COLO829BL (ATCC® CRL-1980™) cell lines were obtained from ATCC in September 2017. A single batch of cells was thawed and cells were expanded and grown according to standard procedures as recommended by ATCC. Cell pellets were split for technology-specific DNA isolation at 33 days (COLO829 & COLO829BL for the ILL and ONT datasets), 35 days (COLO829 for the PB, 10X and BNG datasets) and 23 days (COLO829BL for the PB, 10X and BNG datasets).
4
Isolation of mitochondria from cell lines
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COLO 829 and COLO 829BL (tumor/normal matched) cell lines were purchased from ATCC. DC isolation was performed using the Qproteome kit (Qiagen), and MB isolation with TOM22 antibody bound to MACS magnetic beads was performed using the Mitochondrial Isolation Kit (Miltenyi Biotech), both according to manufacturers’ protocols. MtDNA was extracted from pellet using DNeasy Blood and Tissue kit (Qiagen), eluted using smaller volumes.
5
Comparative Analysis of COLO829 Cell Lines
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COLO829 (ATCC® CRL-1974™) and COLO829BL (ATCC® CRL-1980™) cell lines were obtained from ATCC in September 2017. A single batch of cells was thawed and cells were expanded and grown according to standard procedures as recommended by ATCC. Cell pellets were split for technology-specific DNA isolation at 33 days (COLO829 & COLO829BL for the ILL and ONT datasets), 35 days (COLO829 for the PB, 10X and BNG datasets) and 23 days (COLO829BL for the PB, 10X and BNG datasets).
6
COLO829 and COLO829BL Cell Culture and Freezing
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COLO829 and COLO829BL were obtained from American Type Culture Collection (ATCC), Manassas, VA. Cells were maintained in RPMI 1640 and DMEM media, Life Technologies, Grand Island, NY, respectively. Media was supplemented with 10% heat-inactivated fetal bovine serum (FBS) and Antibiotic-Antimycotic, Life Technologies. Cells were incubated at 37 °C in a humidified 5% CO2 atmosphere and grown to ~80% confluency in T175 tissue culture flasks. COLO829 cells were harvested using trypsin and centrifugation. The non-adherent COLO829BL cells were pelleted by centrifugation. Cell pellets were resuspended in 5 mL of media and aliquots were counted using the Countess Cell Counter, Life Technologies, with the addition of trypan blue to measure viability. Aliquots were made in 15 mL conical tubes ranging from 5 × 106 to 1 × 107 cells per tube and cells were pelleted by centrifugation and immediately flash frozen and stored at −80 C until ready for DNA isolation.
7
Genomic PCR of COLO-829 Cell Lines
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The COLO-829 and COLO-829BL cell lines were purchased from the American Type Culture Collection (ATCC). Primers for genomic PCR were designed using Primer3 (http://biotools.umassmed.edu/bioapps/primer3_www.cgi ). The COLO-829 and COLO-829BL cells were cultured in RPMI 1640(Sigma) supplemented with 10% FBS and 1% penicillin and streptomycin. Genomic DNA was extracted from COLO-829 and COLO-829BL cells using genome DNA kit (Invitrogen) and the PCR was performed using GoTaq DNA Polymerase (Promega). Thermal cycling conditions were one cycles of 95 °C for 2 min, followed by 30 cycles of 95 °C for 30 s, 65°C for 30 s and 72 °C 1 min, followed by a final extension step of 72 °C for 10 min. PCR products were electrophoresed on 1% agarose gels with ethidium bromide, visualized using UV light illumination.
8
Genomic PCR of COLO-829 Cell Lines
9
Tumor Sequencing Protocol for Comprehensive Genomic Analysis
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Tumor specimens were collected from biopsies (needle core, endobronchial ultrasound) or tissue resections (Additional file 2 : Table S2). Solid tumor specimens were snap frozen and liquid biopsies were spun down. Pathology was reviewed and nucleic acids were extracted as described in Pleasance et al. [1 (link)]. Sequencing was performed on either HiSeq 2500 or HiSeq X instruments to target 80X coverage for the tumor samples and 40X coverage for the matched normal samples. The reference cell lines COLO829 and COLO829BL were obtained from American Type Culture Collection (ATCC), Manassas, VA, and sequenced to 80X and 40X coverage, respectively.
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