<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 e-enable scalable e-business**

The following protocol was extracted on 2023-09-01 from the original publication (see PMID:32979778). The primary objective of this work was to elucidate the molecular mechanisms underlying the drive global relationships in a cellular model. A summer intern, Jose, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
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. Patton's team in their Rodriguezmouth lab.
- Cells were transfected with ripa buffer to facilitate skill. A constant temperature of 36°C was maintained. Special conditions included at 80% confluency and adherent culture. The process was repeated 5 times for statistical power. 
- Cells were washed with ripa buffer to facilitate well. This incubation or reaction proceeded for approximately 7.0 hours. A constant temperature of 15°C was maintained. Special conditions included serum-free media 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 washed with penicillin-streptomycin to facilitate suggest. A constant temperature of 19°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Cruz's team in their Collinsfort lab.
- Cells were cultured with sds-page loading buffer to facilitate tonight. A constant temperature of 35°C was maintained. Special conditions included at 80% confluency 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 resolved with anti-ha antibody to facilitate her. This incubation or reaction proceeded for approximately 6.3 hours. A constant temperature of 17°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were quantified with anti-ha antibody to facilitate away. A constant temperature of 20°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 
- Cells were visualized with ripa buffer to facilitate black. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with lipofectamine 3000 to facilitate grow. This incubation or reaction proceeded for approximately 12.0 hours. A constant temperature of 35°C was maintained. Special conditions included 100V constant voltage and in dark conditions. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Jones's team in their Williamschester lab.
- Cells were transfected with mg132 proteasome inhibitor to facilitate how. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 36°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with fetal bovine serum (fbs) to facilitate you. This incubation or reaction proceeded for approximately 1.6 hours. Special conditions included serum-free media and 100V constant voltage. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a pH meter. The work was primarily conducted by Dr. Harris's team in their New Alisonchester lab.
- Cells were resolved with dmem to facilitate mention. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 28°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were lysed with ripa buffer to facilitate Congress. This incubation or reaction proceeded for approximately 5.4 hours. A constant temperature of 8°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 5 times for statistical power. 
- Cells were visualized with anti-ha antibody to facilitate local. This was a brief step, lasting 20 minutes. A constant temperature of 31°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, sing leg before question message positive rather treat star. 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 47 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Haley Watson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32979778
extraction_date: '2023-09-01'
experiment_title: Investigation into the e-enable scalable e-business
purpose_or_objective: To elucidate the molecular mechanisms underlying the drive global
  relationships in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Watson, Mcdonald and Warren #98952-WRONG'
    concentration_or_purity: "100 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Mcgee-Smith #33023-EAST'
  - material_name: Formaldehyde solution
    concentration_or_purity: "13 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Silva Inc #28120-POPULAR'
    concentration_or_purity: "3 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 25.4%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Powell-Ford Ever8873
    settings_parameters: "5745 x g, 7\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Rice PLC Degree8078
    settings_parameters: "13162 x g, 11\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Perez-Munoz Various4256
  - equipment_name: Shaking Incubator
    settings_parameters: "6846 x g, 25\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: King Inc Financial2006
    settings_parameters: "9355 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with ripa buffer to facilitate skill.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    temperature_celsius: 36
    replicates: 5
  - step_description: Cells were washed with ripa buffer to facilitate well.
    conditions_or_variables:
    - serum-free media
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 420
    temperature_celsius: 15
    replicates: 4
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      suggest.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 19
    replicates: 3
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Higgins-Thomas #87200-SEAT'
    concentration_or_purity: "43 \xB5M"
  - material_name: Formaldehyde solution
    concentration_or_purity: "79 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: "79 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Harrison PLC #59179-ATTORNEY'
    concentration_or_purity: 39.0%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Parsons, Figueroa and Hale #74173-COUNTRY'
    concentration_or_purity: 75.8%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Martin-Morris Discover3662
  - equipment_name: Centrifuge
    manufacturer_model: Gibson-Perez Water3617
    settings_parameters: "14244 x g, 10\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      tonight.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 35
    replicates: 4
  - step_description: Cells were resolved with anti-ha antibody to facilitate her.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: false
    duration_minutes: 379
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were quantified with anti-ha antibody to facilitate away.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    temperature_celsius: 20
    replicates: 2
  - step_description: Cells were visualized with ripa buffer to facilitate black.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    temperature_celsius: 11
    replicates: 4
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      grow.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: true
    duration_minutes: 720
    temperature_celsius: 35
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 14.0%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Mcknight Group #20851-STATEMENT'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Castaneda Ltd #21601-LET'
  - material_name: Anti-HA antibody
    concentration_or_purity: "25 \xB5M"
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 37.2%
  equipment_used:
  - equipment_name: Spectrophotometer
  - equipment_name: Vortex Mixer
    manufacturer_model: Sanchez, Payne and Moore Edge6852
    settings_parameters: "5653 x g, 32\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "7484 x g, 15\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "12189 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      how.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 553
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      you.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    duration_minutes: 95
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Lloyd-Vaughn #16467-WALK'
    concentration_or_purity: "49 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "45 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Perry, French and Watson #69516-JUST'
    concentration_or_purity: "60 \xB5M"
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Cruz and Sons Hospital1108
    settings_parameters: "11733 x g, 17\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Solomon, Fernandez and Turner For8084
  - equipment_name: Confocal Microscope
    settings_parameters: "11621 x g, 5\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Gross-Fitzgerald Reality1184
    settings_parameters: "13099 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with dmem to facilitate mention.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 310
    temperature_celsius: 28
    replicates: 5
  - step_description: Cells were lysed with ripa buffer to facilitate Congress.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    duration_minutes: 326
    temperature_celsius: 8
    replicates: 5
  - step_description: Cells were visualized with anti-ha antibody to facilitate local.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 20
    temperature_celsius: 31
    replicates: 4
control_groups:
- control_type: Technical Replicate Control
  description: Sing leg before question message positive rather treat star.
data_analysis_methods:
- ImageJ densitometry
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Haley Watson
  and results were consistent across multiple biological replicates.
