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We narrowed to 18 results for: DME

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Showing: 1 - 18 of 18 results

  1. Cloning Mammalian Cells with the Agarose Method

    Type
    Blog Post
    ...is also placed in the hood. Finally, a sterile 2X DMEM solution (diluted from 10X and containing both Hepes...3 ml of melted agarose are mixed with 3 ml of 2X DMEM in a 50 ml sterile tube (for 60 mm dishes only 1.5...onto the plate. We test the (capped) 1:1 agarose/DMEM solution against a part of the body very sensitive...provide a greater surface area (the amount of agarose/DMEM should be proportionately increased). For some cells...

  4. CRISPR 101: Ribonucleoprotein (RNP) Delivery

    Type
    Blog Post
    ...mediated transfection of protoplasts and biolistic bombardment of immature embryos (Liang et al. 2017).  PEG-mediated..., often show low efficiency, while biolistic bombardment, which can have high efficiency, requires specialized...

  5. Resolute Plasmid Collection

    Type
    Collection
    ...H., Sykacek, P., Reinhardt, J., Müller, A. C., Wiedmer, T., Superti-Fuga, G. (2025). The solute carrier...40355756 (Link opens in a new window) Goldmann, U., Wiedmer, T., Garofoli, A., Sedlyarov, V., Bichler, M., ... a new window) Superti-Furga, G., Lackner, D., Wiedmer, T., Ingles-Prieto, A., Barbosa, B., Girardi, E...window) PMID: 32265506 (Link opens in a new window) Wiedmer, T., Teoh, S.T., Christodoulaki, E., Wolf, G., ...Goldamnn, U., Frommelt, F., Teoh, S. T., Girardi, E., Wiedmer, T., Superti-Furga, G. (2025). The genetic interaction...

  6. Neurodegeneration Plasmid Collection

    Type
    Collection
    ...pnEA-NvM-DmeIF4G_578-650-GB1_Z EIF4G Parkinson's Elisa Izaurralde 148183 pnEA-NvG-DmeIF4G_578-650_Z EIF4G...pnEA-NpM-DmeIF4G_578-675-GB1_AD EIF4G Parkinson's Elisa Izaurralde 148505 pnEA-NpM-DmeIF4G_601-634-GB1_...pnEA-NpM-DmeIF4G_601-660-GB1_AD EIF4G1 Parkinson's Elisa Izaurralde 148507 pnEA-NpM-DmeIF4G_601-675-GB1...pnEA-NpM-DmeIF4G_617-634-GB1_AD EIF4G Parkinson's Elisa Izaurralde 148509 pnEA-NpM-DmeIF4G_617-675-GB1_...Elisa Izaurralde 148512 pnEA-NpM-DmeIF4Ga_617-630-DmThor_64-71-DmeIF4Ga_639-676-GB1_AD EIF4G Parkinson's ...Dementia Walker Jackson 79259 pAc5.1B-lambdaN-HA-DmeIF4G-V5His6 EIF4G1 His, HA, V5 Ac5 Parkinson's Elisa...Parkinson's Denes Hnisz 146392 pAc5.1B-lambdaN-HA-DmeIF4G-trunc-V5His6_G EIF4G His, HA, V5 Ac5 Parkinson'...

  9. AAV Production in HEK293 Cells

    Type
    Protocol
    ... filter will maximize titer . DMEM high glucose, Corning 10-013-CV DMEM, low glucose (1 g/L) glucose, ...HCl, 150 mM NaCl, 2 mM MgCl2) Reagent Preparation DMEM Complete : 10% v/v FBS and 4 mM L-alanyl-L-glutamine...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS ...Store at 4 °C. D1 + 0.1 M sorbitol (optional) : DMEM low glucose + 1% v/v HI-FBS + 0.1 M sorbitol In ...7.5% Sodium Bicarbonate, 7.5 mL 1 M HEPES to 750 mL DMEM + 1 g/L glucose. 1 mg/mL polyethylenimine (PEI) ...Wait ~2 min. Neutralize trypsin by adding 10 mL of DMEM Complete. Pipet back and forth vigorously multiple...from 2 x T-175 flasks. Adjust volume to 300 mL with DMEM complete media and mix. Seed all cells in 1 CS2....

  10. Colony Formation Titering Assay

    Type
    Protocol
    ...Pipet controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine...21008-216 Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat inactivated FBS ...for the colony formation assay. Prepare a batch of DMEM complete containing 10 μg/mL polybrene by diluting... 10-fold serial dilutions of the lentivirus into DMEM complete containing 10 μg/mL polybrene: Dilution...of Lentivirus or Previous Dilution (μL) Volume of DMEM Complete Containing 10 μg/mL Polybrene (μL) Volume...cells as follows: Dilute 7,000 cells into 9.45 mL of DMEM complete containing 10 μg/mL polybrene. Mix well...

  11. Lentivirus Production

    Type
    Protocol
    ...flask in 15 mL DMEM Complete. Wednesday: Plate 1×10 6 cells in a T75 flask in 15 mL DMEM Complete. Friday...container pH meter Stir plate Magnetic Stir Bar Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine...encoding your insert of interest Reagent Preparation DMEM Complete (also known as D10): 10% v/v FBS and 4 ...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS ...Friday: Plate 8×10 5 cells in a T75 flask in 15 mL DMEM Complete. Do not add pen-strep to the media. Use...293T packaging cells at 3.8×10 6 cells per plate in DMEM Complete in 10 cm tissue culture plates. Incubate...15 min at RT. During the incubation, add 10 mL of DMEM Complete to a 15 mL conical and an appropriate amount...

  12. Virus Protocol - Generating Stable Cell Lines

    Type
    Protocol
    ...Pipet controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine...e.g. puromycin, blasticidin) Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS ... that kills all of the cells. Prepare a batch of DMEM complete + 10 µg/mL polybrene by diluting 20 µL ...Prepare a range of dilutions of the lentivirus in DMEM complete + 10 µg/mL polybrene. Note, this is just...well. Dilution Volume of Lentivirus (μL) Volume of DMEM complete + 10 µg/mL polybrene (µL) 0 0 500 1:5 300...Dilute 350,000 cells into a total volume of 7 mL of DMEM complete + 10 µg/mL polybrene. Mix well by pipetting...

  13. General Transfection

    Type
    Protocol
    ...fresh DMEM complete containing 25 µM chloroquine diphosphate and incubate ~5 h For 10 mL of DMEM complete...Pipet controller Hazardous waste container Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine...filter Syringes for filtering Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine...alternative such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS ...Seed HEK293T cells at 3.8x10 6 cells per plate in DMEM complete in 10 cm tissue culture plates. Incubate...aspirate the media. Replace the media with 15 mL of DMEM complete. Incubate the cells 24–48 h before checking...

  14. Lentivirus ddPCR Titration

    Type
    Protocol
    ...Bio-Rad, 10031244 Reagent Preparation DMEM Complete: 500 mL DMEM, high glucose, pyruvate 55 mL heat inactivated...well tissue culture treated dish, VWR, 29442-042 DMEM, high glucose, pyruvate, Corning, 10-013-CV Heat-inactivated...bovine serum 5 mL glutaGRO 50 U/mL benzonase: 15 mL DMEM Complete 3 µL of 250 U/µL benzonase Procedure Transducing...Prepare the following dilution series of virus in DMEM complete: Prepare 7 microcentrifuge tubes with the...well). Leave one well untransduced (add 150 µL of DMEM complete instead of a viral dilution). Remember,...others. Gently add 1.5 mL of 50 U/mL Benzonase in DMEM complete to each well. Incubate at 37 °C for 30 ...µL TrypLE for 1–2 min. Resuspend cells in 500 µL DMEM complete and transfer to a microcentrifuge tube....

  15. Fluorescence Titering Assay

    Type
    Protocol
    ...container Inverted fluorescence microscope Reagents DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine...21008-216 Lentivirus preparation Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS ...cells as follows: Dilute 525,000 cells into 14 mL of DMEM complete. Mix well by pipetting or inverting. Aliquot...to use. Prepare dilutions of the lentivirus into DMEM complete containing 10 μg/mL polybrene. Note, this...Dilution Volume of Lentivirus Stock (μL) Volume of DMEM complete (μL) Volume of 10mg/mL polybrene (μL) 1...

  16. Isolating a Monoclonal Cell Population by Limiting Dilution

    Type
    Protocol
    ...cell counter Reagents Polyclonal stable cell pool DMEM high glucose, Corning 10-013-CV L-alanyl-L-glutamine...dish, Corning 430167 (optional) Reagent Preparation DMEM Complete: 10% v/v FBS and 4 mM L-alanyl-L-glutamine...alternative, such as glutaGRO) To a 500 mL bottle of DMEM high glucose, add 55 mL of heat-inactivated FBS ... dish. Each 10 cm dish should be seeded in 10 mL DMEM complete, which will generate enough conditioned...

  17. Protocol - pLKO.1 – TRC Cloning Vector

    Type
    Protocol
    ...passaged in DMEM + 10% FBS with penicillin/streptomycin, cells should be plated at this step in DMEM + 10% ...Invitrogen: #31985 Dulbecco’s Modified Eagle Medium (DMEM) Invitrogen: #11995 Fetal Bovine Serum (FBS) Invitrogen...incubator. The cells should be 50-80% confluent and in DMEM that does not contain antibiotics. h. Without touching...the transfection reagent. Replace with 5 mL fresh DMEM + 10% FBS + penicillin/streptomycin. Pipette the...
Showing: 1 - 18 of 18 results