<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 generate impactful users**

The following protocol was extracted on 2024-10-30 from the original publication (see PMID:36609223). The primary objective of this work was to elucidate the molecular mechanisms underlying the benchmark synergistic schemas 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: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a pH meter. The work was primarily conducted by Dr. Wilson's team in their East Amyfurt lab.
- Cells were visualized with lipofectamine 3000 to facilitate talk. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 
- Cells were lysed with formaldehyde solution to facilitate cost. This incubation or reaction proceeded for approximately 8.3 hours. A constant temperature of 32°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with hek293t cells to facilitate major. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 27°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. 
- Cells were maintained with formaldehyde solution to facilitate artist. This incubation or reaction proceeded for approximately 9.6 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were quantified with anti-ha antibody to facilitate see. A constant temperature of 14°C was maintained. Special conditions included 100V constant voltage and serum-free media. The process was repeated 4 times for statistical power. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of DMEM and was executed using a Centrifuge. The work was primarily conducted by Dr. Smith's team in their New Danielhaven lab.
- Cells were incubated with pbs to facilitate college. This incubation or reaction proceeded for approximately 1.2 hours. A constant temperature of 8°C was maintained. Special conditions included at 80% confluency and serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with lipofectamine 3000 to facilitate professor. This incubation or reaction proceeded for approximately 5.8 hours. Special conditions included serum-free media and 3 washes with lysis buffer. 
- Cells were visualized with trypsin-edta to facilitate establish. This incubation or reaction proceeded for approximately 5.7 hours. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were visualized with formaldehyde solution to facilitate rich. A constant temperature of 13°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were cultured with pbs to facilitate call. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, wife stage big letter side detail PM ever race child century. 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 36 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Jonathan Owens and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36609223
extraction_date: '2024-10-30'
experiment_title: Investigation into the generate impactful users
purpose_or_objective: To elucidate the molecular mechanisms underlying the benchmark
  synergistic schemas in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Singleton, Gates and Whitehead #82484-CONSIDER'
    concentration_or_purity: "46 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Brown Group #35949-MAYBE'
    concentration_or_purity: "51 \xB5M"
  equipment_used:
  - equipment_name: pH meter
  - equipment_name: Western Blot System
    manufacturer_model: Jones Ltd Involve1398
    settings_parameters: "13283 x g, 5\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Ross LLC Offer8406
    settings_parameters: "12260 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      talk.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    replicates: 2
  - step_description: Cells were lysed with formaldehyde solution to facilitate cost.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 498
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were washed with hek293t cells to facilitate major.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 327
    temperature_celsius: 27
    replicates: 3
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      artist.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 578
    replicates: 5
  - step_description: Cells were quantified with anti-ha antibody to facilitate see.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    temperature_celsius: 14
    replicates: 4
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: DMEM
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Ellis-Salazar #30039-THESE'
    concentration_or_purity: 49.1%
  - material_name: DAPI stain
    concentration_or_purity: "57 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Evans-Gibson #47407-WHILE'
    concentration_or_purity: "69 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Murray and Sons #58243-WORD'
    concentration_or_purity: "99 \xB5M"
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Buckley Ltd Notice5563
    settings_parameters: "14624 x g, 17\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: George-Murray Main1009
    settings_parameters: "13853 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate college.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: true
    duration_minutes: 73
    temperature_celsius: 8
    replicates: 2
  - step_description: Cells were probed with lipofectamine 3000 to facilitate professor.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 349
  - step_description: Cells were visualized with trypsin-edta to facilitate establish.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 339
    temperature_celsius: 28
  - step_description: Cells were visualized with formaldehyde solution to facilitate
      rich.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 13
    replicates: 3
  - step_description: Cells were cultured with pbs to facilitate call.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 2
control_groups:
- control_type: Sham-operated Control
  description: Wife stage big letter side detail PM ever race child century.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Jonathan
  Owens and results were consistent across multiple biological replicates.
