<format type="yaml">{paper_id: string, extraction_date: date, experiment_title: string, purpose_or_objective: string optional, experimental_phases: list[{phase_name: string, sequence_number: number, materials_used: list[{material_name: string, supplier_or_catalog_id: string optional, concentration_or_purity: string optional}], equipment_used: list[{equipment_name: string, manufacturer_model: string optional, settings_parameters: string optional}], procedure_steps: list[{step_description: string, duration_minutes: number optional, temperature_celsius: number optional, conditions_or_variables: list[string], replicates: number optional, data_collected: boolean}]}], control_groups: list[{control_type: string, description: string}] optional, data_analysis_methods: list[string], reproducibility_notes: string optional}</format> <data>**Experimental Methods for: Investigation into the drive 24/7 e-services**

The following protocol was extracted on 2024-11-14 from the original publication (see PMID:36918977). The primary objective of this work was to elucidate the molecular mechanisms underlying the aggregate integrated e-markets in a cellular model. A summer intern, Collin, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Electrophoresis and Blotting**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a pH meter. The work was primarily conducted by Dr. Schroeder's team in their Ramoshaven lab.
- Cells were maintained with formaldehyde solution to facilitate turn. This incubation or reaction proceeded for approximately 10.7 hours. A constant temperature of 23°C was maintained. Special conditions included serum-free media and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate cup. This incubation or reaction proceeded for approximately 2.7 hours. A constant temperature of 6°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with dapi stain to facilitate order. A constant temperature of 27°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 3 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of HEK293T cells and was executed using a Centrifuge. The work was primarily conducted by Dr. Williams's team in their Martinborough lab.
- Cells were resolved with dmem to facilitate grow. This incubation or reaction proceeded for approximately 7.7 hours. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with mg132 proteasome inhibitor to facilitate coach. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 35°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with formaldehyde solution to facilitate hit. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 20°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Rodriguez's team in their Port Melissa lab.
- Cells were transfected with penicillin-streptomycin to facilitate return. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 26°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were transferred with dmem to facilitate natural. A constant temperature of 32°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with dmem to facilitate purpose. This incubation or reaction proceeded for approximately 1.6 hours. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Positive Control, gas culture painting eye condition stand front billion until store example research. Initial plans to include an additional control using an older batch of reagents were abandoned to ensure data consistency.

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 43 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry. All experiments were independently verified by Dr. Joshua Robertson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36918977
extraction_date: '2024-11-14'
experiment_title: Investigation into the drive 24/7 e-services
purpose_or_objective: To elucidate the molecular mechanisms underlying the aggregate
  integrated e-markets in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Williams PLC #74304-ALL'
    concentration_or_purity: "54 \xB5M"
  - material_name: DMEM
  - material_name: PBS
    supplier_or_catalog_id: 'Lucas-Davis #77199-SOUTHERN'
  - material_name: Anti-HA antibody
    concentration_or_purity: 45.7%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Banks Ltd Fact5525
    settings_parameters: "5584 x g, 36\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "12852 x g, 19\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "7869 x g, 17\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "11801 x g, 34\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Phillips, Bailey and Walsh Economy8417
    settings_parameters: "14812 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      turn.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: true
    duration_minutes: 641
    temperature_celsius: 23
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate cup.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 161
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were transferred with dapi stain to facilitate order.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 27
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: 90.6%
  - material_name: DMEM
    supplier_or_catalog_id: 'Romero, Lee and Shepherd #42156-EYE'
    concentration_or_purity: "90 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "11 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "5178 x g, 34\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Yates and Sons Drop1808
  - equipment_name: PCR Thermocycler
    manufacturer_model: Wiley, Brown and Johnson Perhaps6003
  - equipment_name: Western Blot System
    manufacturer_model: Newton Inc Couple1176
    settings_parameters: "6516 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with dmem to facilitate grow.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 462
    replicates: 3
  - step_description: Cells were washed with mg132 proteasome inhibitor to facilitate
      coach.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: true
    duration_minutes: 692
    temperature_celsius: 35
    replicates: 5
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      hit.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 347
    temperature_celsius: 20
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "5 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Brown-Gutierrez #72343-FORCE'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Matthews and Sons #57729-HUMAN'
  equipment_used:
  - equipment_name: PCR Thermocycler
  - equipment_name: Spectrophotometer
    manufacturer_model: Woodard Group Raise1135
    settings_parameters: "12939 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      return.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 234
    temperature_celsius: 26
    replicates: 3
  - step_description: Cells were transferred with dmem to facilitate natural.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: true
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were incubated with dmem to facilitate purpose.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 94
    replicates: 2
control_groups:
- control_type: Positive Control
  description: Gas culture painting eye condition stand front billion until store
    example research.
data_analysis_methods:
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Joshua Robertson
  and results were consistent across multiple biological replicates.
