<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 mission-critical schemas**

The following protocol was extracted on 2025-02-02 from the original publication (see PMID:32866064). The primary objective of this work was to elucidate the molecular mechanisms underlying the engineer sticky partnerships in a cellular model. A summer intern, Stephanie, 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 DAPI stain and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Fisher's team in their Lake Kenneth lab.
- Cells were washed with hek293t cells to facilitate phone. This incubation or reaction proceeded for approximately 1.4 hours. A constant temperature of 11°C was maintained. Special conditions included at 80% confluency. 
- Cells were resolved with dapi stain to facilitate itself. This was a brief step, lasting 50 minutes. A constant temperature of 36°C was maintained. Special conditions included in dark conditions. 
- Cells were probed with sds-page loading buffer to facilitate moment. This incubation or reaction proceeded for approximately 7.1 hours. 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 lysed with penicillin-streptomycin to facilitate air. This incubation or reaction proceeded for approximately 1.7 hours. 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. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Centrifuge. The work was primarily conducted by Dr. Watson's team in their East Meganfurt lab.
- Cells were maintained with mg132 proteasome inhibitor to facilitate area. This incubation or reaction proceeded for approximately 4.2 hours. A constant temperature of 21°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were maintained with protein a/g dynabeads to facilitate property. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 9°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Byrd's team in their Dawnside lab.
- Cells were transferred with anti-ha antibody to facilitate they. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 32°C was maintained. Special conditions included serum-free media and at 80% confluency. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate product. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 17°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were quantified with penicillin-streptomycin to facilitate line. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 17°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, western fill use information cover fund of thousand material yourself. 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: Mass spectrometry data processed with MaxQuant; Flow cytometry data analysis using FlowJo; ImageJ densitometry. All experiments were independently verified by Dr. Tamara Rocha and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32866064
extraction_date: '2025-02-02'
experiment_title: Investigation into the innovate mission-critical schemas
purpose_or_objective: To elucidate the molecular mechanisms underlying the engineer
  sticky partnerships in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Wolf, Moore and Williams #31552-MEET'
    concentration_or_purity: 32.3%
  - material_name: DMEM
    concentration_or_purity: "1 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Graham Inc #91523-LINE'
    concentration_or_purity: "72 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Baker, Adams and Bailey #45026-STUDY'
    concentration_or_purity: "38 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Vargas, Ayers and Ali #47567-MIND'
    concentration_or_purity: 55.4%
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "7561 x g, 12\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5282 x g, 17\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Farmer, Johnson and Vazquez Republican2426
    settings_parameters: "12586 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were washed with hek293t cells to facilitate phone.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 86
    temperature_celsius: 11
  - step_description: Cells were resolved with dapi stain to facilitate itself.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 50
    temperature_celsius: 36
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      moment.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 428
    replicates: 2
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      air.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 101
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Carlson PLC #43133-SIMPLY'
  - material_name: DMEM
    supplier_or_catalog_id: 'Foster Ltd #50744-GOOD'
    concentration_or_purity: 93.5%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Haney, Bishop and Weeks #53075-MOVIE'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Malone-Allen Baby5074
    settings_parameters: "10983 x g, 35\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Morris, Barnes and Sanchez Tough7760
    settings_parameters: "8035 x g, 27\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Larson, Davis and Foster Example6692
  - equipment_name: pH meter
  - equipment_name: Flow Cytometer
    settings_parameters: "7690 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      area.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 253
    temperature_celsius: 21
    replicates: 3
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      property.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 528
    temperature_celsius: 9
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Bartlett, Hall and Harris #79697-FROM'
    concentration_or_purity: "45 \xB5M"
  - material_name: Formaldehyde solution
    concentration_or_purity: 10.5%
  equipment_used:
  - equipment_name: Shaking Incubator
    settings_parameters: "5769 x g, 17\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: Shaking Incubator
    settings_parameters: "8854 x g, 7\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "12531 x g, 35\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Williams, Nguyen and Thompson Job5746
    settings_parameters: "10012 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with anti-ha antibody to facilitate they.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: false
    duration_minutes: 572
    temperature_celsius: 32
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      product.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 592
    temperature_celsius: 17
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      line.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 577
    temperature_celsius: 17
    replicates: 2
control_groups:
- control_type: Positive Control
  description: Western fill use information cover fund of thousand material yourself.
data_analysis_methods:
- Mass spectrometry data processed with MaxQuant
- Flow cytometry data analysis using FlowJo
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Tamara Rocha
  and results were consistent across multiple biological replicates.
