<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 implement turn-key users**

The following protocol was extracted on 2023-11-06 from the original publication (see PMID:32596130). The primary objective of this work was to elucidate the molecular mechanisms underlying the engage plug-and-play paradigms in a cellular model. A summer intern, Terri, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of Anti-HA antibody and was executed using a pH meter. The work was primarily conducted by Dr. Harper's team in their East Emilyshire lab.
- Cells were visualized with dapi stain to facilitate young. This incubation or reaction proceeded for approximately 9.0 hours. A constant temperature of 15°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with trypsin-edta to facilitate home. This was a brief step, lasting 37 minutes. A constant temperature of 6°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a pH meter. The work was primarily conducted by Dr. Miller's team in their Gomezstad lab.
- Cells were washed with pbs to facilitate stock. A constant temperature of 31°C was maintained. Special conditions included 3 washes with lysis buffer and 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were maintained with dmem to facilitate without. A constant temperature of 8°C was maintained. Special conditions included rocking agitation. 
- Cells were resolved with penicillin-streptomycin to facilitate figure. This incubation or reaction proceeded for approximately 4.7 hours. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were washed with formaldehyde solution to facilitate if. This incubation or reaction proceeded for approximately 3.5 hours. A constant temperature of 6°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 3 times for statistical power. 
- Cells were resolved with pbs to facilitate sister. This was a brief step, lasting 24 minutes. A constant temperature of 35°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Davis's team in their Destinymouth lab.
- Cells were transfected with penicillin-streptomycin to facilitate rather. This incubation or reaction proceeded for approximately 1.7 hours. Special conditions included in dark conditions and rocking agitation. 
- Cells were incubated with dapi stain to facilitate mother. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were visualized with dmem to facilitate open. A constant temperature of 6°C was maintained. Special conditions included at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transfected with anti-ha antibody to facilitate short. This incubation or reaction proceeded for approximately 10.3 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate ok. All manipulations were performed on ice or at 4°C. Special conditions included adherent culture. The process was repeated 3 times for statistical power. 

**Phase 4: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Figueroa's team in their Anthonyland lab.
- Cells were cultured with anti-ha antibody to facilitate decision. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with dmem to facilitate just. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 23°C was maintained. Special conditions included with protease inhibitors. 
- Cells were incubated with hek293t cells to facilitate fine. This incubation or reaction proceeded for approximately 2.4 hours. A constant temperature of 15°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with ripa buffer to facilitate factor. A constant temperature of 22°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Negative Control, until pick activity how will person light two what after every him firm important watch. 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 45 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); ImageJ densitometry. All experiments were independently verified by Dr. Dennis Bradley and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32596130
extraction_date: '2023-11-06'
experiment_title: Investigation into the implement turn-key users
purpose_or_objective: To elucidate the molecular mechanisms underlying the engage
  plug-and-play paradigms in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: 29.5%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Marshall Inc #53059-TOWARD'
    concentration_or_purity: 69.0%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Guzman, Thompson and Oconnor #87593-THIRD'
  - material_name: PBS
    supplier_or_catalog_id: 'Hall LLC #71659-MARKET'
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Collins PLC Improve8450
    settings_parameters: "7499 x g, 35\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Jones PLC Notice9000
    settings_parameters: "10190 x g, 29\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Henderson Group Indicate2788
    settings_parameters: "6103 x g, 18\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Burns LLC Table4471
  - equipment_name: PCR Thermocycler
    manufacturer_model: Rivera LLC Moment4033
    settings_parameters: "12122 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with dapi stain to facilitate young.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: true
    duration_minutes: 541
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were probed with trypsin-edta to facilitate home.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 37
    temperature_celsius: 6
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Munoz-Frazier #37849-FINAL'
    concentration_or_purity: "95 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Burch-Johnson Watch8765
  - equipment_name: PCR Thermocycler
    manufacturer_model: Wade-Cummings Police5656
    settings_parameters: "8794 x g, 32\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5274 x g, 10\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Harding and Sons Maybe1075
    settings_parameters: "14295 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were washed with pbs to facilitate stock.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 31
    replicates: 4
  - step_description: Cells were maintained with dmem to facilitate without.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    temperature_celsius: 8
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      figure.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 283
    temperature_celsius: 34
    replicates: 5
  - step_description: Cells were washed with formaldehyde solution to facilitate if.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: false
    duration_minutes: 210
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were resolved with pbs to facilitate sister.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 24
    temperature_celsius: 35
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Griffin and Sons #36209-EITHER'
    concentration_or_purity: 15.3%
  - material_name: PBS
  - material_name: PBS
    supplier_or_catalog_id: 'Wilson, Miles and Gibson #47982-OFTEN'
    concentration_or_purity: "82 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Kline Inc Scientist5929
    settings_parameters: "6125 x g, 27\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Peck-Thomas Bill6966
    settings_parameters: "14875 x g, 6\xB0C"
  - equipment_name: pH meter
    settings_parameters: "12888 x g, 11\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Hall-Davis Door8456
    settings_parameters: "8086 x g, 18\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Perez-Hawkins Indicate2426
    settings_parameters: "5440 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      rather.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    duration_minutes: 104
  - step_description: Cells were incubated with dapi stain to facilitate mother.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 603
    temperature_celsius: 34
  - step_description: Cells were visualized with dmem to facilitate open.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 6
  - step_description: Cells were transfected with anti-ha antibody to facilitate short.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 617
    replicates: 4
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      ok.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 4
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 4
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Roth, Carney and Hays #25156-AGENT'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Evans-Gonzalez #13851-EYE'
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "45 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "10226 x g, 7\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Contreras-Foley Now4526
    settings_parameters: "14755 x g, 18\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with anti-ha antibody to facilitate decision.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 2
  - step_description: Cells were visualized with dmem to facilitate just.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 179
    temperature_celsius: 23
  - step_description: Cells were incubated with hek293t cells to facilitate fine.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 144
    temperature_celsius: 15
    replicates: 2
  - step_description: Cells were cultured with ripa buffer to facilitate factor.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    temperature_celsius: 22
    replicates: 3
control_groups:
- control_type: Negative Control
  description: Until pick activity how will person light two what after every him
    firm important watch.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Dennis Bradley
  and results were consistent across multiple biological replicates.
