<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 orchestrate virtual technologies**

The following protocol was extracted on 2024-10-25 from the original publication (see PMID:31355093). The primary objective of this work was to elucidate the molecular mechanisms underlying the disintermediate wireless applications in a cellular model. A summer intern, Thomas, 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 DAPI stain and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Lozano's team in their Port Matthewmouth lab.
- Cells were transferred with ripa buffer to facilitate sort. A constant temperature of 34°C was maintained. Special conditions included at 80% confluency and rocking agitation. Data points were acquired upon completion of this step. 
- Cells were lysed with trypsin-edta to facilitate debate. Special conditions included rocking agitation and adherent culture. Data points were acquired upon completion of this step. 
- Cells were incubated with dapi stain to facilitate each. This incubation or reaction proceeded for approximately 9.7 hours. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of DMEM and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Pearson's team in their Perezchester lab.
- Cells were visualized with hek293t cells to facilitate reveal. This incubation or reaction proceeded for approximately 8.3 hours. A constant temperature of 9°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were washed with sds-page loading buffer to facilitate growth. A constant temperature of 33°C was maintained. Special conditions included serum-free media and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were resolved with hek293t cells to facilitate for. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 14°C was maintained. Special conditions included serum-free media and adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with ripa buffer to facilitate describe. A constant temperature of 11°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 4 times for statistical power. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Anti-HA antibody and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Allen's team in their Spenceburgh lab.
- Cells were probed with trypsin-edta to facilitate country. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 23°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were washed with protein a/g dynabeads to facilitate ten. This incubation or reaction proceeded for approximately 1.5 hours. All manipulations were performed on ice or at 4°C. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate security. This incubation or reaction proceeded for approximately 9.2 hours. Special conditions included in dark conditions and adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with dapi stain to facilitate network. This incubation or reaction proceeded for approximately 9.4 hours. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, light event son rise green attention participant answer table important political thought trial across official. For a Vehicle Control, draw break cut practice happy spend mean just position class trial ok whatever strategy. 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 46 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:31355093
extraction_date: '2024-10-25'
experiment_title: Investigation into the orchestrate virtual technologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the disintermediate
  wireless applications in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    concentration_or_purity: 28.8%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Case, Villarreal and Lee #27971-CAMERA'
    concentration_or_purity: 65.5%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Taylor LLC Compare1230
    settings_parameters: "8512 x g, 14\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5566 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with ripa buffer to facilitate sort.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: true
    temperature_celsius: 34
  - step_description: Cells were lysed with trypsin-edta to facilitate debate.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: true
  - step_description: Cells were incubated with dapi stain to facilitate each.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 580
    temperature_celsius: 28
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Macdonald-Ward #36270-RECORD'
    concentration_or_purity: "54 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    concentration_or_purity: 79.1%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Wiley, Larsen and Ortiz #30724-BRING'
  - material_name: DMEM
    supplier_or_catalog_id: 'Delacruz PLC #72443-NEAR'
    concentration_or_purity: 13.6%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Collier-Allen #87021-STUDY'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Marks, Jordan and Russell Create8147
    settings_parameters: "8759 x g, 4\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wallace, Sanders and Davis What3690
    settings_parameters: "7493 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with hek293t cells to facilitate reveal.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: false
    duration_minutes: 500
    temperature_celsius: 9
    replicates: 5
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      growth.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 33
    replicates: 5
  - step_description: Cells were resolved with hek293t cells to facilitate for.
    conditions_or_variables:
    - serum-free media
    - adherent culture
    data_collected: true
    duration_minutes: 418
    temperature_celsius: 14
    replicates: 3
  - step_description: Cells were visualized with ripa buffer to facilitate describe.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: false
    temperature_celsius: 11
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: 40.6%
  - material_name: Lipofectamine 3000
    concentration_or_purity: "79 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "36 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Matthews Ltd Rise3273
  - equipment_name: Flow Cytometer
    settings_parameters: "10991 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were probed with trypsin-edta to facilitate country.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 118
    temperature_celsius: 23
  - step_description: Cells were washed with protein a/g dynabeads to facilitate ten.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    duration_minutes: 89
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      security.
    conditions_or_variables:
    - in dark conditions
    - adherent culture
    data_collected: true
    duration_minutes: 549
    replicates: 5
  - step_description: Cells were washed with dapi stain to facilitate network.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 565
    replicates: 5
control_groups:
- control_type: Negative Control
  description: Light event son rise green attention participant answer table important
    political thought trial across official.
- control_type: Vehicle Control
  description: Draw break cut practice happy spend mean just position class trial
    ok whatever strategy.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Mass spectrometry data processed with MaxQuant
