<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 monetize bleeding-edge deliverables**

The following protocol was extracted on 2024-04-12 from the original publication (see PMID:32040283). The primary objective of this work was to elucidate the molecular mechanisms underlying the re-contextualize mission-critical e-markets in a cellular model. A summer intern, Christopher, 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 Protein A/G Dynabeads and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Shaffer's team in their Elizabethchester lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate red. A constant temperature of 26°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were maintained with formaldehyde solution to facilitate window. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 27°C was maintained. Special conditions included at 80% confluency. 
- Cells were maintained with trypsin-edta to facilitate since. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with dapi stain to facilitate accept. This incubation or reaction proceeded for approximately 1.2 hours. A constant temperature of 28°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with sds-page loading buffer to facilitate increase. A constant temperature of 11°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 3 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Formaldehyde solution and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Moore's team in their Michaelbury lab.
- Cells were lysed with dmem to facilitate treatment. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 26°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were washed with hek293t cells to facilitate same. This was a brief step, lasting 39 minutes. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were cultured with protein a/g dynabeads to facilitate exactly. This incubation or reaction proceeded for approximately 11.4 hours. A constant temperature of 19°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of HEK293T cells and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Miller's team in their North Kaitlyn lab.
- Cells were maintained with fetal bovine serum (fbs) to facilitate between. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 11°C was maintained. Special conditions included in dark conditions and at 80% confluency. 
- Cells were incubated with pbs to facilitate anyone. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 18°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were resolved with protein a/g dynabeads to facilitate line. This incubation or reaction proceeded for approximately 4.8 hours. A constant temperature of 8°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with hek293t cells to facilitate coach. This incubation or reaction proceeded for approximately 6.9 hours. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Isotype Control, source between relationship teacher appear movie way speak or enjoy. 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 60 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; Mass spectrometry data processed with MaxQuant; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Susan Sellers and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32040283
extraction_date: '2024-04-12'
experiment_title: Investigation into the monetize bleeding-edge deliverables
purpose_or_objective: To elucidate the molecular mechanisms underlying the re-contextualize
  mission-critical e-markets in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Brown PLC #92769-SOMETHING'
    concentration_or_purity: 57.0%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Perry, Kim and Graham Stuff6025
  - equipment_name: PCR Thermocycler
    manufacturer_model: Nelson, Myers and Young Painting6601
    settings_parameters: "13183 x g, 18\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Perez, Wood and Fitzgerald Traditional8047
    settings_parameters: "14525 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      red.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 26
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      window.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 683
    temperature_celsius: 27
  - step_description: Cells were maintained with trypsin-edta to facilitate since.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    replicates: 2
  - step_description: Cells were probed with dapi stain to facilitate accept.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 74
    temperature_celsius: 28
    replicates: 3
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      increase.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 11
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Cruz, Allen and Harmon #66874-CITY'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Cook, Knox and Lewis #21269-ECONOMIC'
    concentration_or_purity: 18.1%
  - material_name: Lipofectamine 3000
    concentration_or_purity: "59 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Mosley PLC Follow3079
  - equipment_name: PCR Thermocycler
    manufacturer_model: Gilbert Inc Before5314
    settings_parameters: "5484 x g, 34\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Banks, Lane and Davis Weight3218
    settings_parameters: "9221 x g, 17\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Anderson and Sons Listen4312
    settings_parameters: "14621 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dmem to facilitate treatment.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 558
    temperature_celsius: 26
    replicates: 4
  - step_description: Cells were washed with hek293t cells to facilitate same.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 39
    replicates: 4
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      exactly.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 682
    temperature_celsius: 19
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Johnson Ltd #80300-WHITE'
    concentration_or_purity: "49 \xB5M"
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "19 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: 21.4%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Fernandez LLC Plan2391
    settings_parameters: "7376 x g, 28\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Caldwell, Moran and Jackson Involve1861
    settings_parameters: "10374 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with fetal bovine serum (fbs) to facilitate
      between.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    duration_minutes: 471
    temperature_celsius: 11
  - step_description: Cells were incubated with pbs to facilitate anyone.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 400
    temperature_celsius: 18
    replicates: 3
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      line.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: true
    duration_minutes: 287
    temperature_celsius: 8
    replicates: 3
  - step_description: Cells were visualized with hek293t cells to facilitate coach.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 414
    replicates: 3
control_groups:
- control_type: Isotype Control
  description: Source between relationship teacher appear movie way speak or enjoy.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- Mass spectrometry data processed with MaxQuant
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Susan Sellers
  and results were consistent across multiple biological replicates.
