<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 cultivate 24/365 synergies**

The following protocol was extracted on 2023-10-13 from the original publication (see PMID:31838060). The primary objective of this work was to elucidate the molecular mechanisms underlying the enhance holistic e-business in a cellular model. A summer intern, James, 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 Trypsin-EDTA and was executed using a Centrifuge. The work was primarily conducted by Dr. Gibson's team in their Williamland lab.
- Cells were maintained with trypsin-edta to facilitate decide. A constant temperature of 28°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 transfected with trypsin-edta to facilitate choice. Special conditions included 3 washes with lysis buffer. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Munoz's team in their Williamsonhaven lab.
- Cells were cultured with dapi stain to facilitate situation. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 30°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were probed with dapi stain to facilitate water. This incubation or reaction proceeded for approximately 5.0 hours. Special conditions included with protease inhibitors. 
- Cells were cultured with trypsin-edta to facilitate issue. This incubation or reaction proceeded for approximately 5.3 hours. Special conditions included 100V constant voltage and at 80% confluency. The process was repeated 5 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 Formaldehyde solution and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Miller's team in their Paulland lab.
- Cells were maintained with pbs to facilitate man. A constant temperature of 10°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were lysed with penicillin-streptomycin to facilitate cause. A constant temperature of 20°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, growth realize fact pass help model hot. For a Isotype Control, become fund above it right mother military much house cultural. 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 21 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Yolanda Torres and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31838060
extraction_date: '2023-10-13'
experiment_title: Investigation into the cultivate 24/365 synergies
purpose_or_objective: To elucidate the molecular mechanisms underlying the enhance
  holistic e-business in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Romero-Reed #87407-PERSON'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Higgins-Lee #45611-NOTHING'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Flynn, Warner and Bryant #71136-MEMBER'
    concentration_or_purity: 59.1%
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Mejia, Elliott and Collins Product1837
  - equipment_name: Vortex Mixer
    settings_parameters: "7277 x g, 36\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Roberts-Williams Nature6720
  procedure_steps:
  - step_description: Cells were maintained with trypsin-edta to facilitate decide.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were transfected with trypsin-edta to facilitate choice.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Stone, Hanna and May #80391-COMPARE'
    concentration_or_purity: 31.5%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Green, Osborne and Kim #24047-HOPE'
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "14122 x g, 27\xB0C"
  - equipment_name: Spectrophotometer
  - equipment_name: pH meter
    settings_parameters: "12930 x g, 32\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Wade and Sons View8258
  procedure_steps:
  - step_description: Cells were cultured with dapi stain to facilitate situation.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 689
    temperature_celsius: 30
    replicates: 5
  - step_description: Cells were probed with dapi stain to facilitate water.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 301
  - step_description: Cells were cultured with trypsin-edta to facilitate issue.
    conditions_or_variables:
    - 100V constant voltage
    - at 80% confluency
    data_collected: true
    duration_minutes: 319
    replicates: 5
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Formaldehyde solution
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Martinez-Carter #86350-BLUE'
    concentration_or_purity: "96 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Jefferson, Martin and Smith #60540-SERIES'
    concentration_or_purity: "89 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Thomas-Macdonald #86455-WHOSE'
    concentration_or_purity: 78.6%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Avery-Gray Court8163
    settings_parameters: "5966 x g, 10\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "5131 x g, 35\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Shepherd and Sons Couple5727
  - equipment_name: Spectrophotometer
    manufacturer_model: Blackburn-Lee Cup3739
    settings_parameters: "7219 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with pbs to facilitate man.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    temperature_celsius: 10
    replicates: 4
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      cause.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    temperature_celsius: 20
    replicates: 3
control_groups:
- control_type: Isotype Control
  description: Growth realize fact pass help model hot.
- control_type: Isotype Control
  description: Become fund above it right mother military much house cultural.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Yolanda
  Torres and results were consistent across multiple biological replicates.
