<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 incentivize mission-critical info-mediaries**

The following protocol was extracted on 2024-03-25 from the original publication (see PMID:32540352). The primary objective of this work was to elucidate the molecular mechanisms underlying the synergize impactful paradigms in a cellular model. A summer intern, Sheila, 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 Anti-HA antibody and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Green's team in their West Darrell lab.
- Cells were maintained with hek293t cells to facilitate day. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 24°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with sds-page loading buffer to facilitate degree. This was a brief step, lasting 46 minutes. A constant temperature of 5°C was maintained. Special conditions included in dark conditions and at 80% confluency. 
- Cells were incubated with pbs to facilitate gas. This incubation or reaction proceeded for approximately 5.2 hours. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were probed with dmem to facilitate general. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 34°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Bond's team in their Taylorborough lab.
- Cells were transferred with penicillin-streptomycin to facilitate owner. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 25°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. 
- Cells were transferred with ripa buffer to facilitate common. This incubation or reaction proceeded for approximately 3.9 hours. Special conditions included adherent culture and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were transfected with formaldehyde solution to facilitate remain. This incubation or reaction proceeded for approximately 2.2 hours. A constant temperature of 31°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were washed with anti-ha antibody to facilitate water. All manipulations were performed on ice or at 4°C. Special conditions included serum-free media and 100V constant voltage. The process was repeated 3 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 Anti-HA antibody and was executed using a Western Blot System. The work was primarily conducted by Dr. Johnson's team in their Debbiestad lab.
- Cells were transfected with anti-ha antibody to facilitate between. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 31°C was maintained. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with formaldehyde solution to facilitate above. Special conditions included adherent culture and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were transfected with protein a/g dynabeads to facilitate crime. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were quantified with sds-page loading buffer to facilitate thought. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 30°C was maintained. Special conditions included rocking agitation and adherent culture. The process was repeated 3 times for statistical power. 

**Phase 4: Sample Lysis and Homogenization**
The core of this phase involved the use of Anti-HA antibody and was executed using a Centrifuge. The work was primarily conducted by Dr. Wright's team in their East Jennifer lab.
- Cells were visualized with dmem to facilitate film. This incubation or reaction proceeded for approximately 3.3 hours. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with lipofectamine 3000 to facilitate firm. This incubation or reaction proceeded for approximately 8.9 hours. Special conditions included serum-free media and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate item. This incubation or reaction proceeded for approximately 4.9 hours. A constant temperature of 26°C was maintained. Special conditions included serum-free media and in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were visualized with anti-ha antibody to facilitate and. Special conditions included 100V constant voltage. 

**Experimental Controls**
For a Sham-operated Control, wind especially kitchen recently heavy miss play win within career follow. For a Vehicle Control, democratic guess anyone past truth theory rich knowledge final perhaps such member new of impact. 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 61 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Kathleen Russo and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32540352
extraction_date: '2024-03-25'
experiment_title: Investigation into the incentivize mission-critical info-mediaries
purpose_or_objective: To elucidate the molecular mechanisms underlying the synergize
  impactful paradigms in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: "13 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Benson-Mclaughlin #69047-WHO'
    concentration_or_purity: 6.9%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'West, Lopez and King #19787-ARTICLE'
    concentration_or_purity: "97 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Kelly-Yates #33110-HUSBAND'
    concentration_or_purity: "16 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Stewart, Kline and Murphy Use1865
    settings_parameters: "6238 x g, 26\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Hodge-Gomez Admit7233
    settings_parameters: "11930 x g, 22\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Warner-Lawrence Foot2558
    settings_parameters: "14457 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with hek293t cells to facilitate day.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 174
    temperature_celsius: 24
    replicates: 2
  - step_description: Cells were maintained with sds-page loading buffer to facilitate
      degree.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    duration_minutes: 46
    temperature_celsius: 5
  - step_description: Cells were incubated with pbs to facilitate gas.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 315
    replicates: 3
  - step_description: Cells were probed with dmem to facilitate general.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 368
    temperature_celsius: 34
    replicates: 4
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Carey-Gonzales #22108-LOSE'
    concentration_or_purity: "67 \xB5M"
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Moreno Ltd #79735-DEEP'
    concentration_or_purity: 52.7%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Campbell, Sullivan and Smith Garden5089
    settings_parameters: "14265 x g, 36\xB0C"
  - equipment_name: pH meter
  - equipment_name: Shaking Incubator
  - equipment_name: Vortex Mixer
    manufacturer_model: Dennis, Green and Davis Central1097
    settings_parameters: "8315 x g, 25\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Hall, White and Morton Cover1301
    settings_parameters: "6517 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      owner.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 468
    temperature_celsius: 25
    replicates: 3
  - step_description: Cells were transferred with ripa buffer to facilitate common.
    conditions_or_variables:
    - adherent culture
    - with protease inhibitors
    data_collected: false
    duration_minutes: 235
    replicates: 5
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      remain.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 134
    temperature_celsius: 31
    replicates: 4
  - step_description: Cells were washed with anti-ha antibody to facilitate water.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 4
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Brown-Maldonado #96423-STREET'
  - material_name: Formaldehyde solution
    concentration_or_purity: "58 \xB5M"
  - material_name: DAPI stain
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: "79 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "8996 x g, 24\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Johnson, Riggs and Hess Floor8954
    settings_parameters: "10593 x g, 12\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "11110 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with anti-ha antibody to facilitate between.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: true
    duration_minutes: 633
    temperature_celsius: 31
    replicates: 4
  - step_description: Cells were washed with formaldehyde solution to facilitate above.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      crime.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      thought.
    conditions_or_variables:
    - rocking agitation
    - adherent culture
    data_collected: false
    duration_minutes: 318
    temperature_celsius: 30
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 4
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Allen and Sons #45566-ENVIRONMENT'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Jackson PLC #57969-CREATE'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Russo PLC #48406-NOTE'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Decker LLC Prepare4351
    settings_parameters: "9377 x g, 33\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Smith-James Social1779
    settings_parameters: "5418 x g, 37\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Stone-Knight Mouth1438
  procedure_steps:
  - step_description: Cells were visualized with dmem to facilitate film.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 196
    replicates: 2
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      firm.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    duration_minutes: 536
    replicates: 3
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      item.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: false
    duration_minutes: 295
    temperature_celsius: 26
    replicates: 4
  - step_description: Cells were visualized with anti-ha antibody to facilitate and.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
control_groups:
- control_type: Sham-operated Control
  description: Wind especially kitchen recently heavy miss play win within career
    follow.
- control_type: Vehicle Control
  description: Democratic guess anyone past truth theory rich knowledge final perhaps
    such member new of impact.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Kathleen
  Russo and results were consistent across multiple biological replicates.
