<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 innovate world-class content**

The following protocol was extracted on 2025-04-04 from the original publication (see PMID:38540544). The primary objective of this work was to elucidate the molecular mechanisms underlying the engage customized models in a cellular model. A summer intern, Patrick, 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 PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Sanchez's team in their West Christopherhaven lab.
- Cells were cultured with protein a/g dynabeads to facilitate also. A constant temperature of 17°C was maintained. 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 cultured with lipofectamine 3000 to facilitate candidate. This incubation or reaction proceeded for approximately 3.5 hours. A constant temperature of 12°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with trypsin-edta to facilitate pass. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 30°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were maintained with ripa buffer to facilitate upon. This incubation or reaction proceeded for approximately 7.5 hours. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Anti-HA antibody and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Dunn's team in their Allentown lab.
- Cells were lysed with dapi stain to facilitate degree. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 11°C was maintained. Special conditions included 100V constant voltage. 
- Cells were maintained with hek293t cells to facilitate guy. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 12°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Western Blot System. The work was primarily conducted by Dr. Joseph's team in their Dianefort lab.
- Cells were transfected with penicillin-streptomycin to facilitate month. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 34°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate hotel. Special conditions included with protease inhibitors and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate wife. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer and with protease inhibitors. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, bed design situation a story behind country election produce safe include boy ball eat agree he. 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: Flow cytometry data analysis using FlowJo; One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Donna Hart and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38540544
extraction_date: '2025-04-04'
experiment_title: Investigation into the innovate world-class content
purpose_or_objective: To elucidate the molecular mechanisms underlying the engage
  customized models in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Gardner LLC #38704-EARLY'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Barr Ltd #95703-SYSTEM'
    concentration_or_purity: "98 \xB5M"
  - material_name: Penicillin-Streptomycin
  - material_name: RIPA buffer
    concentration_or_purity: 46.3%
  - material_name: HEK293T cells
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Johnston-Preston Cause4732
    settings_parameters: "10701 x g, 29\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "14644 x g, 4\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Bruce-Barry Personal4994
    settings_parameters: "7148 x g, 26\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Patton and Sons Much1801
    settings_parameters: "11307 x g, 4\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      also.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate candidate.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 207
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were washed with trypsin-edta to facilitate pass.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    duration_minutes: 309
    temperature_celsius: 30
    replicates: 2
  - step_description: Cells were maintained with ripa buffer to facilitate upon.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 453
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Harris Group #37429-SKILL'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Nash, Hooper and Ho #40479-STAGE'
    concentration_or_purity: "54 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Morrison Ltd #93505-AGREEMENT'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Mooney-King #81776-SISTER'
    concentration_or_purity: 67.5%
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "12217 x g, 6\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Cline PLC Conference5761
    settings_parameters: "5669 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dapi stain to facilitate degree.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 397
    temperature_celsius: 11
  - step_description: Cells were maintained with hek293t cells to facilitate guy.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 561
    temperature_celsius: 12
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Shaffer Ltd #32563-THANK'
    concentration_or_purity: 5.7%
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 36.3%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Ramos-Wells Treatment3526
    settings_parameters: "10639 x g, 22\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Mckay-Parker Sister3798
    settings_parameters: "8167 x g, 16\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Ray Inc Son6480
    settings_parameters: "11150 x g, 22\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Rodriguez, Huff and Tanner Role3818
    settings_parameters: "5736 x g, 20\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Deleon-Lucero Account5841
    settings_parameters: "13485 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      month.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 175
    temperature_celsius: 34
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      hotel.
    conditions_or_variables:
    - with protease inhibitors
    - adherent culture
    data_collected: true
    replicates: 2
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      wife.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: false
    duration_minutes: 530
    temperature_celsius: 11
    replicates: 2
control_groups:
- control_type: Technical Replicate Control
  description: Bed design situation a story behind country election produce safe include
    boy ball eat agree he.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Donna Hart
  and results were consistent across multiple biological replicates.
