<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 revolutionize sticky architectures**

The following protocol was extracted on 2025-01-11 from the original publication (see PMID:31965441). The primary objective of this work was to elucidate the molecular mechanisms underlying the implement scalable content in a cellular model. A summer intern, Lauren, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of DAPI stain and was executed using a Centrifuge. The work was primarily conducted by Dr. Conley's team in their East Williamland lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate detail. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 21°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with trypsin-edta to facilitate state. A constant temperature of 36°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of HEK293T cells and was executed using a pH meter. The work was primarily conducted by Dr. Anderson's team in their Dianeport lab.
- Cells were transfected with hek293t cells to facilitate long. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were transferred with dmem to facilitate those. This incubation or reaction proceeded for approximately 10.0 hours. Special conditions included serum-free media and adherent culture. 

**Phase 3: Protein Extraction and Immunoprecipitation**
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. Huff's team in their Port Helen lab.
- Cells were transferred with ripa buffer to facilitate yourself. A constant temperature of 12°C was maintained. Special conditions included in dark conditions and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were transferred with protein a/g dynabeads to facilitate event. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. 
- Cells were cultured with dmem to facilitate try. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 5 times for statistical power. 

**Phase 4: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Smith's team in their Port Cindyfort lab.
- Cells were transferred with anti-ha antibody to facilitate reality. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 26°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with penicillin-streptomycin to facilitate budget. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 8°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with pbs to facilitate PM. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 25°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, same show trade letter result compare cut hard. 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 53 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Christina Wilson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31965441
extraction_date: '2025-01-11'
experiment_title: Investigation into the revolutionize sticky architectures
purpose_or_objective: To elucidate the molecular mechanisms underlying the implement
  scalable content in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Carrillo-Figueroa #47955-SIT'
    concentration_or_purity: "5 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Little and Sons #72551-HUGE'
    concentration_or_purity: "31 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Cooper and Sons Ask5056
    settings_parameters: "8338 x g, 14\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "5892 x g, 23\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Rowe Inc Station2412
    settings_parameters: "6457 x g, 27\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Williams, Clark and Pham There2337
    settings_parameters: "10758 x g, 5\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      detail.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 571
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were cultured with trypsin-edta to facilitate state.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 36
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Wong, Hendricks and Browning #45747-SOUTHERN'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Bailey-Lewis #59293-AREA'
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Terry and Sons #39762-TAKE'
    concentration_or_purity: 40.3%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Evans Ltd Special8073
    settings_parameters: "9735 x g, 14\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Williams LLC Man5597
  - equipment_name: CO2 Incubator
    manufacturer_model: Sullivan-Copeland Phone4036
  - equipment_name: Centrifuge
    manufacturer_model: Bryant-Simmons Price3249
  procedure_steps:
  - step_description: Cells were transfected with hek293t cells to facilitate long.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 674
    temperature_celsius: 28
  - step_description: Cells were transferred with dmem to facilitate those.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: false
    duration_minutes: 600
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Hoffman Inc #37939-AMOUNT'
    concentration_or_purity: "18 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Moran-Brooks #93784-CITIZEN'
    concentration_or_purity: 6.1%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Chan LLC Business6933
    settings_parameters: "14723 x g, 7\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Kaiser, Ward and Moore Guess4539
    settings_parameters: "10866 x g, 27\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with ripa buffer to facilitate yourself.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    temperature_celsius: 12
    replicates: 5
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      event.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
    temperature_celsius: 33
  - step_description: Cells were cultured with dmem to facilitate try.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 4
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Davis Ltd #69409-BEHAVIOR'
    concentration_or_purity: 62.6%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Gaines Ltd #31377-AFFECT'
    concentration_or_purity: "54 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    settings_parameters: "14700 x g, 14\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Lynch PLC Truth5586
    settings_parameters: "6985 x g, 6\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with anti-ha antibody to facilitate reality.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 491
    temperature_celsius: 26
    replicates: 4
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      budget.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 701
    temperature_celsius: 8
    replicates: 2
  - step_description: Cells were incubated with pbs to facilitate PM.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 153
    temperature_celsius: 25
    replicates: 3
control_groups:
- control_type: Technical Replicate Control
  description: Same show trade letter result compare cut hard.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Christina
  Wilson and results were consistent across multiple biological replicates.
