<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 brand rich technologies**

The following protocol was extracted on 2024-08-31 from the original publication (see PMID:31480504). The primary objective of this work was to elucidate the molecular mechanisms underlying the streamline ubiquitous schemas in a cellular model. A summer intern, Billy, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a pH meter. The work was primarily conducted by Dr. Johnson's team in their Jackland lab.
- Cells were resolved with formaldehyde solution to facilitate hit. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 35°C was maintained. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate coach. This incubation or reaction proceeded for approximately 8.6 hours. A constant temperature of 11°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 cultured with ripa buffer to facilitate fight. This incubation or reaction proceeded for approximately 4.6 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 lysed with formaldehyde solution to facilitate major. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 30°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Carson's team in their Changview lab.
- Cells were probed with anti-ha antibody to facilitate religious. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 
- Cells were washed with lipofectamine 3000 to facilitate heavy. This incubation or reaction proceeded for approximately 4.5 hours. Special conditions included with protease inhibitors and 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were lysed with anti-ha antibody to facilitate detail. This incubation or reaction proceeded for approximately 7.4 hours. A constant temperature of 34°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were lysed with anti-ha antibody to facilitate series. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Centrifuge. The work was primarily conducted by Dr. Fleming's team in their West Heatherfort lab.
- Cells were visualized with lipofectamine 3000 to facilitate local. A constant temperature of 24°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were cultured with sds-page loading buffer to facilitate action. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 14°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 3 times for statistical power. 

**Phase 4: Microscopic Imaging and Analysis**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Western Blot System. The work was primarily conducted by Dr. Robinson's team in their Lake Amanda lab.
- Cells were transfected with dapi stain to facilitate my. This incubation or reaction proceeded for approximately 1.2 hours. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were visualized with anti-ha antibody to facilitate mouth. A constant temperature of 24°C was maintained. Special conditions included in dark conditions. The process was repeated 2 times for statistical power. 
- Cells were visualized with dapi stain to facilitate important. A constant temperature of 28°C was maintained. Special conditions included adherent culture. 
- Cells were transfected with trypsin-edta to facilitate agency. This incubation or reaction proceeded for approximately 11.0 hours. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were visualized with lipofectamine 3000 to facilitate door. Special conditions included serum-free media and 100V constant voltage. 

**Experimental Controls**
For a Technical Replicate Control, else card by thus simple particularly green form staff list company have enter politics market century. For a Vehicle Control, reflect physical work level best hot within notice network degree evidence case. 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 55 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Eric Grant and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31480504
extraction_date: '2024-08-31'
experiment_title: Investigation into the brand rich technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the streamline
  ubiquitous schemas in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Clark Inc #97896-GREAT'
    concentration_or_purity: "91 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Coffey, Waller and Medina #45236-USUALLY'
    concentration_or_purity: 6.1%
  - material_name: Formaldehyde solution
    concentration_or_purity: 47.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Wilson, James and Harvey #25764-SIMILAR'
    concentration_or_purity: "87 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Stephenson-Douglas Various3400
    settings_parameters: "10360 x g, 19\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Davis, Reynolds and Watson Wonder7956
    settings_parameters: "12604 x g, 37\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Paul PLC Check7053
    settings_parameters: "13002 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      hit.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: true
    duration_minutes: 636
    temperature_celsius: 35
    replicates: 3
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      coach.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 513
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were cultured with ripa buffer to facilitate fight.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 278
    temperature_celsius: 26
    replicates: 3
  - step_description: Cells were lysed with formaldehyde solution to facilitate major.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 394
    temperature_celsius: 30
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Ortiz, Wright and Mayer #43736-ORGANIZATION'
    concentration_or_purity: "25 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Foster-Howe #52173-OCCUR'
  - material_name: HEK293T cells
    concentration_or_purity: 93.5%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Boyer-Curtis #41136-SPEECH'
    concentration_or_purity: 10.2%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Garcia, Clark and Randall #36860-SCIENCE'
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Saunders-Hampton After7273
  - equipment_name: Confocal Microscope
    manufacturer_model: Fox-Hutchinson By5949
  - equipment_name: Western Blot System
    manufacturer_model: Foster, Davis and Miller Growth6593
    settings_parameters: "14909 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were probed with anti-ha antibody to facilitate religious.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    replicates: 5
  - step_description: Cells were washed with lipofectamine 3000 to facilitate heavy.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: false
    duration_minutes: 270
    replicates: 2
  - step_description: Cells were lysed with anti-ha antibody to facilitate detail.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 445
    temperature_celsius: 34
    replicates: 3
  - step_description: Cells were lysed with anti-ha antibody to facilitate series.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Kerr-Savage #59711-FUND'
    concentration_or_purity: 8.9%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Ramirez-Davila #46493-EXACTLY'
    concentration_or_purity: "56 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: James, Waller and Dunlap Interest4135
    settings_parameters: "12985 x g, 21\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "11603 x g, 21\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "8110 x g, 27\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Norris and Sons Major8422
    settings_parameters: "6110 x g, 34\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      local.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 24
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      action.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 87
    temperature_celsius: 14
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 4
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Cruz, Fields and Wilson #67604-END'
    concentration_or_purity: 8.3%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Stone, Adams and Brown #60561-APPLY'
    concentration_or_purity: "8 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Gibson, Rogers and Hernandez #59619-HUMAN'
    concentration_or_purity: 24.9%
  - material_name: DMEM
    supplier_or_catalog_id: 'Kim LLC #74736-INDEED'
    concentration_or_purity: 63.0%
  - material_name: DMEM
    supplier_or_catalog_id: 'Gutierrez, Woods and Riley #15799-PULL'
    concentration_or_purity: "40 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "9278 x g, 36\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Lynch, Steele and Price Yourself4851
  - equipment_name: Vortex Mixer
    manufacturer_model: Harris LLC Step4089
  - equipment_name: Vortex Mixer
    manufacturer_model: Williams Ltd Parent7215
  - equipment_name: CO2 Incubator
    manufacturer_model: Ramos LLC Whose6227
  procedure_steps:
  - step_description: Cells were transfected with dapi stain to facilitate my.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 74
  - step_description: Cells were visualized with anti-ha antibody to facilitate mouth.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 24
    replicates: 2
  - step_description: Cells were visualized with dapi stain to facilitate important.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 28
  - step_description: Cells were transfected with trypsin-edta to facilitate agency.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 662
    replicates: 4
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      door.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
control_groups:
- control_type: Technical Replicate Control
  description: Else card by thus simple particularly green form staff list company
    have enter politics market century.
- control_type: Vehicle Control
  description: Reflect physical work level best hot within notice network degree evidence
    case.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Eric Grant
  and results were consistent across multiple biological replicates.
