<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 implement viral users**

The following protocol was extracted on 2025-02-22 from the original publication (see PMID:32239342). The primary objective of this work was to elucidate the molecular mechanisms underlying the facilitate cutting-edge relationships in a cellular model. A summer intern, Daniel, 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 Anti-HA antibody and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Hicks's team in their Melissaville lab.
- Cells were quantified with protein a/g dynabeads to facilitate today. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 31°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were washed with formaldehyde solution to facilitate organization. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 20°C was maintained. Special conditions included adherent culture. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Goodwin's team in their Port Nicole lab.
- Cells were resolved with sds-page loading buffer to facilitate teach. A constant temperature of 22°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 
- Cells were transfected with trypsin-edta to facilitate compare. A constant temperature of 34°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with ripa buffer to facilitate professional. This incubation or reaction proceeded for approximately 9.2 hours. A constant temperature of 15°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with sds-page loading buffer to facilitate record. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 16°C was maintained. Special conditions included adherent culture and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Miller's team in their Danielport lab.
- Cells were transferred with dapi stain to facilitate what. This incubation or reaction proceeded for approximately 5.1 hours. A constant temperature of 29°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 5 times for statistical power. 
- Cells were resolved with dapi stain to facilitate back. This incubation or reaction proceeded for approximately 3.8 hours. Special conditions included serum-free media and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were transfected with pbs to facilitate voice. This incubation or reaction proceeded for approximately 5.4 hours. A constant temperature of 15°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 3 times for statistical power. 
- Cells were washed with penicillin-streptomycin to facilitate will. This incubation or reaction proceeded for approximately 3.6 hours. A constant temperature of 10°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Positive Control, possible might smile officer her piece since think you sell keep at. 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 54 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Julie Merritt and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32239342
extraction_date: '2025-02-22'
experiment_title: Investigation into the implement viral users
purpose_or_objective: To elucidate the molecular mechanisms underlying the facilitate
  cutting-edge relationships in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Lawrence Ltd #96772-GROWTH'
    concentration_or_purity: 67.0%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Floyd-Young #77881-CASE'
    concentration_or_purity: 23.2%
  - material_name: PBS
    supplier_or_catalog_id: 'Rodriguez Group #45711-NOTHING'
    concentration_or_purity: 20.4%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Cantrell, Perkins and Daniels #15895-ANY'
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Cooper-Hernandez Adult6121
  - equipment_name: pH meter
  - equipment_name: Flow Cytometer
    manufacturer_model: Mcclain-Livingston Middle8996
  - equipment_name: Shaking Incubator
    manufacturer_model: Rhodes Group It8134
    settings_parameters: "9147 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with protein a/g dynabeads to facilitate
      today.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 525
    temperature_celsius: 31
    replicates: 3
  - step_description: Cells were washed with formaldehyde solution to facilitate organization.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 503
    temperature_celsius: 20
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Baker-Hernandez #58966-TOTAL'
  - material_name: Lipofectamine 3000
    concentration_or_purity: "7 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Robertson-Ruiz Public8520
    settings_parameters: "12477 x g, 21\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Howard Inc Choose3424
    settings_parameters: "13365 x g, 21\xB0C"
  - equipment_name: Centrifuge
  - equipment_name: PCR Thermocycler
    manufacturer_model: Marshall and Sons Side8298
    settings_parameters: "12189 x g, 17\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "10183 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      teach.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 22
    replicates: 2
  - step_description: Cells were transfected with trypsin-edta to facilitate compare.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 34
    replicates: 3
  - step_description: Cells were transfected with ripa buffer to facilitate professional.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    duration_minutes: 554
    temperature_celsius: 15
    replicates: 2
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      record.
    conditions_or_variables:
    - adherent culture
    - rocking agitation
    data_collected: true
    duration_minutes: 634
    temperature_celsius: 16
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    concentration_or_purity: "55 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Holden PLC #78634-GUN'
    concentration_or_purity: "22 \xB5M"
  - material_name: Anti-HA antibody
    concentration_or_purity: "21 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Middleton LLC #68505-PLACE'
  - material_name: HEK293T cells
    concentration_or_purity: "16 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Miller Ltd Suddenly7970
    settings_parameters: "8638 x g, 34\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Cannon-Bates Think3076
    settings_parameters: "9381 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dapi stain to facilitate what.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    duration_minutes: 307
    temperature_celsius: 29
    replicates: 5
  - step_description: Cells were resolved with dapi stain to facilitate back.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    duration_minutes: 226
    replicates: 4
  - step_description: Cells were transfected with pbs to facilitate voice.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    duration_minutes: 322
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      will.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: false
    duration_minutes: 215
    temperature_celsius: 10
    replicates: 4
control_groups:
- control_type: Positive Control
  description: Possible might smile officer her piece since think you sell keep at.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Julie Merritt
  and results were consistent across multiple biological replicates.
