Circlegrow media

Manufactured by MP Biomedicals

Sourced in United States

CircleGrow media is a sterile, pre-prepared culture medium designed for the growth and maintenance of various cell lines in circular culture vessels. The media provides the necessary nutrients and growth factors required to support the proliferation of cells in a closed, circular environment.

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Lab products found in correlation

Xbai, by Thermo Fisher Scientific (1 mentions) Top10 competent cell, by Thermo Fisher Scientific (1 mentions) Bamhi, by Thermo Fisher Scientific (1 mentions) Polyethyleneimine pei, by Merck Group (1 mentions) Bseri, by New England Biolabs (1 mentions) Pcr purification kit, by GeneAll (1 mentions) Endofree plasmid maxi kit, by Qiagen (1 mentions) Nebuilder hifi dna assembly kit, by New England Biolabs (1 mentions) Clonejet pcr cloning kit, by Thermo Fisher Scientific (1 mentions) Hispur cobalt resin, by Thermo Fisher Scientific (1 mentions) Ampicillin, by Avantor (1 mentions) Copper chloride, by Merck Group (1 mentions) Isopropyl β d 1 thiogalactopyranoside (iptg), by Biosynth (1 mentions) Fetal bovine serum (fbs), by Merck Group (1 mentions) Sodium chloride (nacl), by Merck Group (1 mentions) Mda mb 231 human breast cancer cells, by American Type Culture Collection (1 mentions) 4t1 murine breast cancer cell, by American Type Culture Collection (1 mentions)

4 protocols using circlegrow media

Bacterial Expression Vector Preparation

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Example 1

The pET-21a vector and BL21 (DE3) E. coli cells were obtained from Novagen Inc. (Madison, Wisconsin, US). Top10 competent cells were obtained from Invitrogen (Carlsbad, California, US). Oligonucleotides were synthesized chemically at Cosmo Gene Tech (Seoul, South Korea). The FastAP thermosensitive alkaline phosphatase and restriction endonuclease including BamHI and XbaI were purchased from Fermentas (Ontario, Canada). Other restriction endonuclease including BseRI and AcuI and all other restriction enzymes were obtained from New England Biolabs (Ipswich, Massachusetts, US). DNA miniprep, gel extraction and PCR purification kits were obtained from Geneall Biotechnology (Seoul, South Korea). Dyne Agarose High was obtained from Dyne Bio, Inc. (Seongnam, South Korea). All the Top10 cells were grown in TB DRY media obtained from MO Bio Laboratories, Inc. (Carlsbad. California, US). All the BL21 (DE3) cells were grown in CircleGrow media obtained from MP Biomedicals (Solon, Ohio, US). Ready Gel (Tris-HCl 2-20%) as a precast gel was purchased from Bio-Rad (Hercules, California, US). Phosphate-buffered saline (PBS, pH 7.4), ampicillin and polyethyleneimine (PEI) were obtained from Sigma-Aldrich (St Louis, Missouri).

US11827679B2. Self-assembled nanostructures of elastin- and resilin-based block copolypeptides with stimuli responsiveness and resilience for drug delivery system, tissue engineering and regenerative medicine and methods of preparing the same (2023-11-28). Industry-University Cooperation Foundation Hanyang University Erica Campus [KR]. Inventors: Dong Woo Lim [KR], Aamna Basheer [KR], Jae Sang Lee [KR], Min Jung Kang [KR].

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CRISPR-Mediated Gene Editing in Mammalian Cells

Cited 1 time

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The CHOPCHOP website (https://chopchop.rc.fas.harvard.edu/) was utilized to design a high-performance sgRNA targeting the human AAVS1 locus. Based on our prior experience, we preferentially chose a sgRNA with a G at the 5′ end which initiates U6-promoter-mediated transcription [20 (link)]. The sgRNA used in this study was sgAAVS1b.
Both the Cas9 and sgRNA plasmids were constructed with a NEBuilder HiFi DNA Assembly Kit (New England Biolabs). Multiple colonies were chosen for Sanger sequencing (MCLAB) to identify the correct clones using the primers U6-F, EF1-F, and wpre-R. Correct clones were grown in CircleGrow Media (MP Biomedicals) and DNA plasmids were purified using Endo-Free Plasmid Maxi Kits (QiaGen).
The TPO-E2A-GFP and GFP donor plasmids used in this study were generated with a CloneJET PCR Cloning Kit (Thermo Scientific). Multiple colonies were chosen for Sanger sequencing (MCLAB) to identify the correct clones using the primers pJET1.2-F and pJET1.2-R. Correct clones were cultured, and DNA plasmids were purified, as described previously [20 (link)] (See also Additional file 1: Additional Information for details). All of the primer sequences are listed in Additional file 1: Table S1.

Zhang L., Liu C., Wang H., Wu D., Su P., Wang M., Guo J., Zhao S., Dong S., Zhou W., Arakaki C., Zhang X, & Zhou J. (2018). Thrombopoietin knock-in augments platelet generation from human embryonic stem cells. Stem Cell Research & Therapy, 9, 194.

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FECH and PGRMC1 Co-Expression in E. coli

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Expression of his-tagged wild-type FECH, FECH variants, and non-tagged PGRMC1 in E. coli was carried out by growth in Circlegrow media (MP Biomedicals, Santa Ana, CA) for 18–20 hours at 30°C. Cells were harvested by centrifugation at 5,000 x g for 10 min, resuspended in solubilization buffer (50 mM Tris-MOPS, pH 8.0, 100 mM KCl, 1% sodium cholate) and sonicated three times on ice for 30 seconds. The resulting lysate was then centrifuged at 100,000xg for 20 minutes and the supernatant reserved. Supernatant from non-tagged PGRMC1 was then mixed with the his-tagged wild-type and variant FECH supernatant and loaded onto HisPur Cobalt Resin (Thermo Fisher Scientific). The column was then washed with wash buffer (50 mM Tris-MOPS pH 8.1, 100 mM KCl, 1% sodium cholate, 15 mM imidazole) and subsequently eluted using elution buffer (50 mM Tris-MOPS pH 8.1, 100 mM KCl, 1% sodium cholate, 250 mM imidazole). Presence of PGRMC1 and relative amounts of PGRMC1 and FECH, both wild-type and variants, were analyzed by SDS-PAGE and immunoblots.

Piel RB I.I.I., Shiferaw M.T., Vashisht A.A., Marcero J., Praissman J., Phillips J.D., Wohlschlegel J.A, & Medlock A.E. (2016). A Novel Role for Progesterone Receptor Membrane Component 1 (PGRMC1): A Partner and Regulator of Ferrochelatase. Biochemistry, 55(37), 5204-5217.

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Protein Expression and Labeling

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Synthetic oligonucleotides encoding monomer genes of V₇, V₃G₃A₃, and AP1-V₅ were obtained from Macrogen Inc. Seoul. All the restriction enzymes were purchased from New England Biolab. The competent BL21 (DE3) E. coli cells and DH5α were procured from Invitrogen, Carlsbad, CA, USA. Circle grow media (MP Biomedicals, CA, USA), 100 μg mL-1 of ampicillin (AMRESCO, LLC, OH, USA) and 1 mM IPTG (Carbosynth Limited, Berkshire, UK) were used for protein expression. Sodium chloride and copper chloride were acquired from Sigma Aldrich, St. Louis, MO, USA. Flamma 675 Vinylsulfone was obtained from BioActs, Incheon, Korea. Sulfo-SMCC (succinimide 4-[N-maleimidomethyl] cyclohexane carboxylate) and other chemicals are acquired from Sigma Aldrich. 4T1 murine breast cancer cells and MDA MB231 human breast cancer cells were obtained from the American Type Culture Collection (ATCC). 4T1 cells were grown in RPMI-1640 (Hyclone), and MDA MB231 cells were grown in Dulbecco's modified Eagle's medium (DMEM) containing 10 % fetal bovine serum (Sigma Aldrich), 100 U/mL of penicillin, and 100 μg/mL of streptomycin (Hyclone). Cells were maintained at 37 °C containing 5% CO₂.

Sarangthem V., Seo B.Y., Yi A., Lee Y.J., Cheon S.H., Kim S.K., Singh T.D., Lee B.H, & Park R.W. (2020). Effects of molecular weight and structural conformation of multivalent-based elastin-like polypeptides on tumor accumulation and tissue biodistribution. Nanotheranostics, 4(2), 57-70.

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