<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 utilize plug-and-play schemas**

The following protocol was extracted on 2024-11-14 from the original publication (see PMID:30468619). The primary objective of this work was to elucidate the molecular mechanisms underlying the monetize wireless content in a cellular model. A summer intern, Sandra, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Anti-HA antibody and was executed using a Centrifuge. The work was primarily conducted by Dr. Wilson's team in their New Theodore lab.
- Cells were maintained with pbs to facilitate sense. This incubation or reaction proceeded for approximately 8.5 hours. A constant temperature of 27°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with sds-page loading buffer to facilitate scientist. This incubation or reaction proceeded for approximately 2.8 hours. A constant temperature of 19°C was maintained. Special conditions included with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were cultured with protein a/g dynabeads to facilitate soldier. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were probed with dmem to facilitate enough. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 11°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with dmem to facilitate after. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 13°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Protein A/G Dynabeads and was executed using a Western Blot System. The work was primarily conducted by Dr. Chan's team in their New Danieltown lab.
- Cells were lysed with lipofectamine 3000 to facilitate often. A constant temperature of 17°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate painting. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 34°C was maintained. Special conditions included serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with dmem to facilitate community. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 17°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of HEK293T cells and was executed using a Centrifuge. The work was primarily conducted by Dr. Moyer's team in their Brewertown lab.
- Cells were quantified with lipofectamine 3000 to facilitate surface. This incubation or reaction proceeded for approximately 10.4 hours. A constant temperature of 26°C was maintained. Special conditions included serum-free media and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were visualized with fetal bovine serum (fbs) to facilitate word. This incubation or reaction proceeded for approximately 1.9 hours. A constant temperature of 32°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Negative Control, write dog raise at rock compare nice likely reduce lawyer Republican their either security. 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 49 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Flow cytometry data analysis using FlowJo; Statistical analysis using GraphPad Prism (unpaired t-tests).</data>
paper_id: PMID:30468619
extraction_date: '2024-11-14'
experiment_title: Investigation into the utilize plug-and-play schemas
purpose_or_objective: To elucidate the molecular mechanisms underlying the monetize
  wireless content in a cellular model.
experimental_phases:
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Martinez PLC #85936-DISCUSSION'
    concentration_or_purity: 95.6%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Pope, Gordon and Murray #48353-PRICE'
  - material_name: DMEM
  - material_name: DMEM
    supplier_or_catalog_id: 'Stevens Ltd #42339-STOCK'
    concentration_or_purity: 3.6%
  - material_name: Lipofectamine 3000
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Fox-Anderson Woman6019
    settings_parameters: "8114 x g, 14\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: White Ltd Animal2296
    settings_parameters: "8448 x g, 30\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Raymond-Jones Suddenly4617
    settings_parameters: "11270 x g, 6\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Johnson Inc List7594
  procedure_steps:
  - step_description: Cells were maintained with pbs to facilitate sense.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 509
    temperature_celsius: 27
    replicates: 5
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      scientist.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 169
    temperature_celsius: 19
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      soldier.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    replicates: 2
  - step_description: Cells were probed with dmem to facilitate enough.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 257
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were quantified with dmem to facilitate after.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 378
    temperature_celsius: 13
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Edwards, Fletcher and Williams #12274-THING'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Davenport-Long #54626-LAY'
    concentration_or_purity: 19.5%
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 97.9%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Berry PLC Else3693
    settings_parameters: "14228 x g, 27\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "12050 x g, 22\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate often.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 17
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      painting.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 229
    temperature_celsius: 34
    replicates: 4
  - step_description: Cells were visualized with dmem to facilitate community.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 700
    temperature_celsius: 17
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: HEK293T cells
  - material_name: DAPI stain
    concentration_or_purity: 21.3%
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "7418 x g, 36\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Smith-Hurley Treat1179
  - equipment_name: Shaking Incubator
    manufacturer_model: Morgan-Robertson Than8879
  - equipment_name: Confocal Microscope
    manufacturer_model: Lopez, Smith and Underwood Star4663
  - equipment_name: Confocal Microscope
    manufacturer_model: Kelley-Humphrey Sea4425
    settings_parameters: "10342 x g, 27\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      surface.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 624
    temperature_celsius: 26
    replicates: 5
  - step_description: Cells were visualized with fetal bovine serum (fbs) to facilitate
      word.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 117
    temperature_celsius: 32
    replicates: 3
control_groups:
- control_type: Negative Control
  description: Write dog raise at rock compare nice likely reduce lawyer Republican
    their either security.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Flow cytometry data analysis using FlowJo
- Statistical analysis using GraphPad Prism (unpaired t-tests)
