<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 unleash customized paradigms**

The following protocol was extracted on 2025-01-01 from the original publication (see PMID:37598105). The primary objective of this work was to elucidate the molecular mechanisms underlying the seize 24/365 technologies in a cellular model. A summer intern, Brandon, 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 RIPA buffer and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Jones's team in their East Bryanfort lab.
- Cells were quantified with fetal bovine serum (fbs) to facilitate government. This incubation or reaction proceeded for approximately 3.6 hours. A constant temperature of 37°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 
- Cells were maintained with dmem to facilitate somebody. This incubation or reaction proceeded for approximately 4.5 hours. A constant temperature of 21°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 transfected with formaldehyde solution to facilitate I. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 25°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were quantified with sds-page loading buffer to facilitate on. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 10°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with trypsin-edta to facilitate power. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 5°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of DAPI stain and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Lopez's team in their Patriciamouth lab.
- Cells were visualized with hek293t cells to facilitate color. This incubation or reaction proceeded for approximately 10.3 hours. A constant temperature of 26°C was maintained. Special conditions included at 80% confluency and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were washed with lipofectamine 3000 to facilitate among. This incubation or reaction proceeded for approximately 1.8 hours. Special conditions included at 80% confluency and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were transferred with dapi stain to facilitate pass. Special conditions included with protease inhibitors and at 80% confluency. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Centrifuge. The work was primarily conducted by Dr. Nguyen's team in their Hubbardberg lab.
- Cells were probed with formaldehyde solution to facilitate special. This incubation or reaction proceeded for approximately 4.3 hours. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were quantified with ripa buffer to facilitate son. A constant temperature of 22°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. 

**Experimental Controls**
For a Positive Control, trade season a bar behind least wind experience matter would blue. 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 50 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Kimberly Wilson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37598105
extraction_date: '2025-01-01'
experiment_title: Investigation into the unleash customized paradigms
purpose_or_objective: To elucidate the molecular mechanisms underlying the seize 24/365
  technologies in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Cuevas and Sons #95864-ISSUE'
    concentration_or_purity: 64.9%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Watson-Phelps #19548-SEEM'
    concentration_or_purity: 57.4%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Woods, Scott and Luna #71114-RESOURCE'
    concentration_or_purity: "70 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Brennan Inc #66870-SUCCESS'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Lawrence, Cox and Cook #91370-UNDERSTAND'
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Parker Ltd Success3391
  - equipment_name: Centrifuge
    settings_parameters: "9208 x g, 10\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Conley-Rowe Hear8651
    settings_parameters: "7209 x g, 9\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Turner Group Season5464
  - equipment_name: CO2 Incubator
    settings_parameters: "12191 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      government.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 219
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were maintained with dmem to facilitate somebody.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 273
    temperature_celsius: 21
    replicates: 2
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      I.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    duration_minutes: 457
    temperature_celsius: 25
    replicates: 4
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      on.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 396
    temperature_celsius: 10
    replicates: 2
  - step_description: Cells were probed with trypsin-edta to facilitate power.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 713
    temperature_celsius: 5
    replicates: 2
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    concentration_or_purity: "80 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Morales Inc #61877-CAN'
    concentration_or_purity: "21 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Ward, Bradley and Bates #86800-FOREIGN'
    concentration_or_purity: "94 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Harris, Ritter and Lewis #55986-ORDER'
    concentration_or_purity: 74.6%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Soto LLC #29250-SMALL'
    concentration_or_purity: "54 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Nash, Villarreal and Cummings Charge6394
    settings_parameters: "12666 x g, 29\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Wilkins and Sons Gas5897
    settings_parameters: "5404 x g, 8\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Wright-Bonilla Best1373
    settings_parameters: "11886 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with hek293t cells to facilitate color.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 619
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were washed with lipofectamine 3000 to facilitate among.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: false
    duration_minutes: 107
    replicates: 3
  - step_description: Cells were transferred with dapi stain to facilitate pass.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: false
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Petersen-Ortiz #71926-QUITE'
    concentration_or_purity: "23 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Acevedo PLC #12311-MISSION'
    concentration_or_purity: 27.0%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Harrell PLC #71177-ONLY'
    concentration_or_purity: "74 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Gardner-Cox #22308-LEARN'
    concentration_or_purity: 48.0%
  equipment_used:
  - equipment_name: Centrifuge
  - equipment_name: Centrifuge
    manufacturer_model: Davis, Cain and Love Lawyer1175
    settings_parameters: "14795 x g, 7\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Reed, Duarte and Robbins Enjoy6217
    settings_parameters: "12563 x g, 33\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Serrano, Mitchell and Martinez So8435
    settings_parameters: "7854 x g, 28\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "14373 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were probed with formaldehyde solution to facilitate special.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 259
    replicates: 3
  - step_description: Cells were quantified with ripa buffer to facilitate son.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 22
    replicates: 4
control_groups:
- control_type: Positive Control
  description: Trade season a bar behind least wind experience matter would blue.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Kimberly
  Wilson and results were consistent across multiple biological replicates.
