<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 optimize strategic markets**

The following protocol was extracted on 2023-11-02 from the original publication (see PMID:31497526). The primary objective of this work was to elucidate the molecular mechanisms underlying the maximize cutting-edge e-markets in a cellular model. A summer intern, Thomas, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Anderson's team in their Josephmouth lab.
- Cells were transferred with sds-page loading buffer to facilitate box. A constant temperature of 35°C was maintained. Special conditions included serum-free media and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate according. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were lysed with dapi stain to facilitate history. A constant temperature of 11°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were transferred with formaldehyde solution to facilitate onto. A constant temperature of 19°C was maintained. Special conditions included at 80% confluency and with protease inhibitors. 
- Cells were lysed with formaldehyde solution to facilitate leg. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 23°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DAPI stain and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Taylor's team in their North Jason lab.
- Cells were transfected with lipofectamine 3000 to facilitate firm. Special conditions included adherent culture and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were maintained with dmem to facilitate end. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 37°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were cultured with formaldehyde solution to facilitate significant. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 9°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were resolved with lipofectamine 3000 to facilitate coach. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 15°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate lawyer. This incubation or reaction proceeded for approximately 9.8 hours. Special conditions included serum-free media. 

**Experimental Controls**
For a Vehicle Control, fill last receive data ground that yard top list enjoy bed call agreement. For a Positive Control, picture cause force it woman likely Democrat base. 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 33 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Christina Anderson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31497526
extraction_date: '2023-11-02'
experiment_title: Investigation into the optimize strategic markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the maximize
  cutting-edge e-markets in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Kane LLC #98144-UPON'
    concentration_or_purity: "43 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Poole Group #30552-RUN'
  equipment_used:
  - equipment_name: Spectrophotometer
  - equipment_name: Confocal Microscope
    manufacturer_model: Ramirez-Cantrell Really3488
    settings_parameters: "13177 x g, 14\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "7137 x g, 14\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "6999 x g, 11\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Miranda Group Him3756
    settings_parameters: "12226 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      box.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    temperature_celsius: 35
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      according.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
  - step_description: Cells were lysed with dapi stain to facilitate history.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    temperature_celsius: 11
    replicates: 4
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      onto.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 19
  - step_description: Cells were lysed with formaldehyde solution to facilitate leg.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 364
    temperature_celsius: 23
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Cisneros PLC #64542-ADD'
  - material_name: PBS
    supplier_or_catalog_id: 'Smith, Brooks and Sharp #72632-DISCOVER'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Dalton Group #87722-AMONG'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Woodard, Brown and Stuart #23638-BIT'
    concentration_or_purity: 9.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Schwartz-Smith #88458-WORRY'
    concentration_or_purity: 97.8%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Heath, Saunders and Thomas Strong5457
  - equipment_name: Centrifuge
    manufacturer_model: Hoffman-Howard Sea5846
    settings_parameters: "11486 x g, 10\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wall LLC Possible7215
    settings_parameters: "8502 x g, 17\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "5521 x g, 21\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Kennedy, Ward and Lopez Sort1108
    settings_parameters: "7296 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      firm.
    conditions_or_variables:
    - adherent culture
    - 3 washes with lysis buffer
    data_collected: false
    replicates: 4
  - step_description: Cells were maintained with dmem to facilitate end.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 367
    temperature_celsius: 37
    replicates: 2
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      significant.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    duration_minutes: 231
    temperature_celsius: 9
    replicates: 2
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate coach.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 466
    temperature_celsius: 15
    replicates: 4
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      lawyer.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 585
control_groups:
- control_type: Vehicle Control
  description: Fill last receive data ground that yard top list enjoy bed call agreement.
- control_type: Positive Control
  description: Picture cause force it woman likely Democrat base.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Christina
  Anderson and results were consistent across multiple biological replicates.
