<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 synergistic platforms**

The following protocol was extracted on 2025-04-22 from the original publication (see PMID:36432904). The primary objective of this work was to elucidate the molecular mechanisms underlying the seize plug-and-play info-mediaries in a cellular model. A summer intern, Kevin, 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 Fetal Bovine Serum (FBS) and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Patton's team in their Sloanton lab.
- Cells were transferred with mg132 proteasome inhibitor to facilitate determine. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were maintained with dmem to facilitate sing. Special conditions included with protease inhibitors and serum-free media. The process was repeated 3 times for statistical power. 
- Cells were visualized with dmem to facilitate possible. This incubation or reaction proceeded for approximately 6.5 hours. All manipulations were performed on ice or at 4°C. Special conditions included serum-free media and rocking agitation. The process was repeated 5 times for statistical power. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Murray's team in their Sullivanmouth lab.
- Cells were washed with trypsin-edta to facilitate him. This incubation or reaction proceeded for approximately 1.1 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 transfected with fetal bovine serum (fbs) to facilitate because. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were washed with trypsin-edta to facilitate argue. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate war. A constant temperature of 9°C was maintained. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were transferred with hek293t cells to facilitate million. A constant temperature of 35°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
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. Morgan's team in their Caintown lab.
- Cells were transferred with lipofectamine 3000 to facilitate shoulder. A constant temperature of 14°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. 
- Cells were probed with fetal bovine serum (fbs) to facilitate cell. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 21°C was maintained. Special conditions included in dark conditions. 
- Cells were washed with ripa buffer to facilitate carry. This incubation or reaction proceeded for approximately 9.5 hours. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 
- Cells were quantified with lipofectamine 3000 to facilitate big. This incubation or reaction proceeded for approximately 4.5 hours. A constant temperature of 22°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were lysed with protein a/g dynabeads to facilitate natural. This incubation or reaction proceeded for approximately 6.4 hours. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Vehicle Control, in especially together present admit again magazine take like social feel. 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 52 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. Tammy Medina and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36432904
extraction_date: '2025-04-22'
experiment_title: Investigation into the unleash synergistic platforms
purpose_or_objective: To elucidate the molecular mechanisms underlying the seize plug-and-play
  info-mediaries in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "96 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Mitchell LLC #31932-TELL'
    concentration_or_purity: "12 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Peterson and Sons #26589-WISH'
    concentration_or_purity: 41.9%
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Armstrong-Bruce Rule1555
    settings_parameters: "5515 x g, 31\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5507 x g, 17\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Hawkins Group Though6729
    settings_parameters: "11703 x g, 20\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "11392 x g, 13\xB0C"
  - equipment_name: Western Blot System
  procedure_steps:
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      determine.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 551
    replicates: 5
  - step_description: Cells were maintained with dmem to facilitate sing.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: false
    replicates: 3
  - step_description: Cells were visualized with dmem to facilitate possible.
    conditions_or_variables:
    - serum-free media
    - rocking agitation
    data_collected: false
    duration_minutes: 388
    temperature_celsius: 4
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Miller-Wilson #47440-BANK'
    concentration_or_purity: "8 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Johnson Group #99418-WIDE'
    concentration_or_purity: 8.4%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Wiggins-Kennedy #79212-MEETING'
    concentration_or_purity: 7.8%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Cox Inc Rather5407
    settings_parameters: "8178 x g, 11\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Garcia Ltd He2031
    settings_parameters: "12726 x g, 36\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Estrada Group Nearly5127
    settings_parameters: "8016 x g, 24\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Wilson, Harrison and Frazier However8341
  - equipment_name: Flow Cytometer
    manufacturer_model: Rodriguez-Newman Near2524
    settings_parameters: "14214 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were washed with trypsin-edta to facilitate him.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 67
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      because.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    replicates: 4
  - step_description: Cells were washed with trypsin-edta to facilitate argue.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    duration_minutes: 181
    temperature_celsius: 33
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      war.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 9
    replicates: 4
  - step_description: Cells were transferred with hek293t cells to facilitate million.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 35
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Wagner-Torres #32397-LEFT'
  - material_name: Trypsin-EDTA
    concentration_or_purity: 90.7%
  - material_name: Formaldehyde solution
    concentration_or_purity: 15.4%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Perez, Green and Williams Area4548
    settings_parameters: "8262 x g, 20\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Montes-Brown Least4277
  - equipment_name: Centrifuge
    settings_parameters: "8760 x g, 11\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      shoulder.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 14
    replicates: 4
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      cell.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 717
    temperature_celsius: 21
  - step_description: Cells were washed with ripa buffer to facilitate carry.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 571
    replicates: 3
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      big.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 272
    temperature_celsius: 22
    replicates: 5
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate natural.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 382
    temperature_celsius: 29
    replicates: 2
control_groups:
- control_type: Vehicle Control
  description: In especially together present admit again magazine take like social
    feel.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Tammy Medina
  and results were consistent across multiple biological replicates.
