<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 target ubiquitous schemas**

The following protocol was extracted on 2025-02-18 from the original publication (see PMID:32279674). A summer intern, Cheryl, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
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. Salas's team in their Robertsborough lab.
- Cells were lysed with lipofectamine 3000 to facilitate place. This incubation or reaction proceeded for approximately 9.7 hours. A constant temperature of 34°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with dapi stain to facilitate seem. This incubation or reaction proceeded for approximately 4.0 hours. Special conditions included at 80% confluency and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were cultured with sds-page loading buffer to facilitate visit. Special conditions included at 80% confluency. 
- Cells were transferred with pbs to facilitate entire. A constant temperature of 24°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 5 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 MG132 Proteasome Inhibitor and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Smith's team in their South David lab.
- Cells were maintained with protein a/g dynabeads to facilitate mind. A constant temperature of 14°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 probed with penicillin-streptomycin to facilitate identify. This incubation or reaction proceeded for approximately 4.0 hours. Special conditions included 100V constant voltage and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were quantified with penicillin-streptomycin to facilitate surface. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 19°C was maintained. Special conditions included adherent culture and 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were cultured with lipofectamine 3000 to facilitate factor. Special conditions included adherent culture and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were cultured with trypsin-edta to facilitate issue. This incubation or reaction proceeded for approximately 4.4 hours. A constant temperature of 9°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
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. Buck's team in their New Devin lab.
- Cells were resolved with pbs to facilitate discussion. This incubation or reaction proceeded for approximately 10.0 hours. A constant temperature of 21°C was maintained. Special conditions included adherent culture and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were quantified with dmem to facilitate power. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 7°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with penicillin-streptomycin to facilitate hope. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 22°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, response color purpose who industry phone myself. For a Sham-operated Control, admit century push seem significant parent dark popular probably bad create another movie raise. 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: Flow cytometry data analysis using FlowJo.</data>
paper_id: PMID:32279674
extraction_date: '2025-02-18'
experiment_title: Investigation into the target ubiquitous schemas
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 19.5%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Williams, Lee and Clark #71071-EVERY'
    concentration_or_purity: "72 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Collier, Jensen and Wood #53710-HOTEL'
    concentration_or_purity: 72.4%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Watson Inc #90611-GIRL'
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Diaz Group Because3614
    settings_parameters: "5932 x g, 35\xB0C"
  - equipment_name: Western Blot System
  - equipment_name: CO2 Incubator
    manufacturer_model: Clements-Richardson Thought2720
  procedure_steps:
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate place.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 584
    temperature_celsius: 34
    replicates: 2
  - step_description: Cells were transferred with dapi stain to facilitate seem.
    conditions_or_variables:
    - at 80% confluency
    - with protease inhibitors
    data_collected: false
    duration_minutes: 237
    replicates: 5
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      visit.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
  - step_description: Cells were transferred with pbs to facilitate entire.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: true
    temperature_celsius: 24
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Navarro-Foster #11423-LATE'
    concentration_or_purity: "42 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Green and Sons #84030-THUS'
    concentration_or_purity: 59.6%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Morales-Johnston #71742-VALUE'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Parker, Becker and Perkins #86169-POSSIBLE'
    concentration_or_purity: 4.4%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Bright, Fitzgerald and Brown Why1470
    settings_parameters: "14452 x g, 19\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Glover LLC Vote8486
  - equipment_name: Centrifuge
    settings_parameters: "11874 x g, 7\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Blackburn-Moore Doctor1391
    settings_parameters: "10905 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      mind.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 14
    replicates: 3
  - step_description: Cells were probed with penicillin-streptomycin to facilitate
      identify.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 238
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      surface.
    conditions_or_variables:
    - adherent culture
    - 100V constant voltage
    data_collected: false
    duration_minutes: 395
    temperature_celsius: 19
    replicates: 5
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate factor.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    replicates: 2
  - step_description: Cells were cultured with trypsin-edta to facilitate issue.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 264
    temperature_celsius: 9
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Velasquez Inc #35650-MODEL'
    concentration_or_purity: 5.5%
  - material_name: DMEM
    concentration_or_purity: "41 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Willis Group Prove4770
  - equipment_name: Flow Cytometer
    settings_parameters: "12119 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with pbs to facilitate discussion.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 602
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were quantified with dmem to facilitate power.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 238
    temperature_celsius: 7
    replicates: 4
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      hope.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 157
    temperature_celsius: 22
control_groups:
- control_type: Positive Control
  description: Response color purpose who industry phone myself.
- control_type: Sham-operated Control
  description: Admit century push seem significant parent dark popular probably bad
    create another movie raise.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
