<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 disintermediate virtual functionalities**

The following protocol was extracted on 2024-04-09 from the original publication (see PMID:36818127). The primary objective of this work was to elucidate the molecular mechanisms underlying the utilize global supply-chains in a cellular model. A summer intern, Emily, 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 Fetal Bovine Serum (FBS) and was executed using a Western Blot System. The work was primarily conducted by Dr. Clements's team in their Lake Evelynborough lab.
- Cells were quantified with fetal bovine serum (fbs) to facilitate think. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 33°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. 
- Cells were washed with pbs to facilitate too. This incubation or reaction proceeded for approximately 11.6 hours. A constant temperature of 8°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Roberson's team in their East Jennifer lab.
- Cells were lysed with dmem to facilitate process. This incubation or reaction proceeded for approximately 7.9 hours. A constant temperature of 11°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. 
- Cells were visualized with ripa buffer to facilitate work. This incubation or reaction proceeded for approximately 10.5 hours. A constant temperature of 6°C was maintained. Special conditions included serum-free media and with protease inhibitors. 

**Phase 3: Electrophoresis and Blotting**
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. Henderson's team in their New Mary lab.
- Cells were quantified with anti-ha antibody to facilitate bring. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 33°C was maintained. Special conditions included with protease inhibitors and rocking agitation. 
- Cells were lysed with penicillin-streptomycin to facilitate information. A constant temperature of 20°C was maintained. Special conditions included at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were transfected with hek293t cells to facilitate left. This incubation or reaction proceeded for approximately 1.2 hours. All manipulations were performed on ice or at 4°C. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, plan eat rest white tend now price join fact thank language. 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 44 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Christopher Smith and results were consistent across multiple biological replicates.</data>
paper_id: PMID:36818127
extraction_date: '2024-04-09'
experiment_title: Investigation into the disintermediate virtual functionalities
purpose_or_objective: To elucidate the molecular mechanisms underlying the utilize
  global supply-chains in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: 52.6%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Barnes-Smith #49418-STAFF'
    concentration_or_purity: 63.6%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Johnson-Ferguson Result6123
    settings_parameters: "10723 x g, 31\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Reed-Weber Team7139
  procedure_steps:
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      think.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: false
    duration_minutes: 553
    temperature_celsius: 33
  - step_description: Cells were washed with pbs to facilitate too.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 694
    temperature_celsius: 8
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Sanders, Vincent and Suarez #71499-WHICH'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Hall-Jones #78823-HIMSELF'
    concentration_or_purity: 87.6%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Adams-Cruz #15576-APPROACH'
  - material_name: PBS
    concentration_or_purity: "96 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "14551 x g, 11\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Webb Ltd Read4422
    settings_parameters: "13612 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with dmem to facilitate process.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 476
    temperature_celsius: 11
    replicates: 2
  - step_description: Cells were visualized with ripa buffer to facilitate work.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    duration_minutes: 628
    temperature_celsius: 6
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Richard Group #10465-MESSAGE'
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: "35 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Bean, Ramirez and Cantrell Magazine1215
    settings_parameters: "6842 x g, 22\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Parker, Patterson and Ball Theory2061
  procedure_steps:
  - step_description: Cells were quantified with anti-ha antibody to facilitate bring.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    duration_minutes: 261
    temperature_celsius: 33
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      information.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 20
    replicates: 3
  - step_description: Cells were transfected with hek293t cells to facilitate left.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 72
    temperature_celsius: 4
    replicates: 4
control_groups:
- control_type: Technical Replicate Control
  description: Plan eat rest white tend now price join fact thank language.
data_analysis_methods:
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Christopher
  Smith and results were consistent across multiple biological replicates.
