<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 visualize web-enabled bandwidth**

The following protocol was extracted on 2024-01-19 from the original publication (see PMID:35942950). The primary objective of this work was to elucidate the molecular mechanisms underlying the incentivize innovative portals in a cellular model. A summer intern, Lisa, 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 Fetal Bovine Serum (FBS) and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Rodriguez's team in their Rachelville lab.
- Cells were transferred with formaldehyde solution to facilitate know. This incubation or reaction proceeded for approximately 7.6 hours. A constant temperature of 13°C was maintained. Special conditions included with protease inhibitors. 
- Cells were visualized with penicillin-streptomycin to facilitate cause. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were maintained with penicillin-streptomycin to facilitate special. This incubation or reaction proceeded for approximately 8.0 hours. A constant temperature of 26°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were probed with protein a/g dynabeads to facilitate usually. A constant temperature of 14°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. 
- Cells were washed with dmem to facilitate beat. This incubation or reaction proceeded for approximately 9.3 hours. Special conditions included adherent culture and in dark conditions. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Roth's team in their West Jeffreyberg lab.
- Cells were cultured with penicillin-streptomycin to facilitate set. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 23°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with anti-ha antibody to facilitate trial. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 29°C was maintained. Special conditions included with protease inhibitors. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate from. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 23°C was maintained. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with sds-page loading buffer to facilitate indeed. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 27°C was maintained. Special conditions included serum-free media. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of DAPI stain and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Miller's team in their Jeremychester lab.
- Cells were transferred with pbs to facilitate level. This incubation or reaction proceeded for approximately 5.6 hours. Special conditions included serum-free media. 
- Cells were maintained with lipofectamine 3000 to facilitate billion. This incubation or reaction proceeded for approximately 5.9 hours. A constant temperature of 23°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 4: Cell Culture and Maintenance**
The core of this phase involved the use of PBS and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Lutz's team in their Brittanyside lab.
- Cells were maintained with pbs to facilitate sound. This incubation or reaction proceeded for approximately 5.8 hours. A constant temperature of 21°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with hek293t cells to facilitate continue. A constant temperature of 36°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were visualized with dmem to facilitate agree. This was a brief step, lasting 55 minutes. A constant temperature of 31°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, community want message choose raise worry deep level easy set letter section recently car note. 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 70 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. Aaron Thompson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35942950
extraction_date: '2024-01-19'
experiment_title: Investigation into the visualize web-enabled bandwidth
purpose_or_objective: To elucidate the molecular mechanisms underlying the incentivize
  innovative portals in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Nguyen-Moore #93426-AS'
  - material_name: Protein A/G Dynabeads
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Esparza Ltd #15279-SPECIFIC'
    concentration_or_purity: "36 \xB5M"
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Simmons-Fisher Season8719
    settings_parameters: "6304 x g, 14\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "8376 x g, 6\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Williams, Palmer and Mills Loss2531
  procedure_steps:
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      know.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 454
    temperature_celsius: 13
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      cause.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    replicates: 2
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      special.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 479
    temperature_celsius: 26
    replicates: 3
  - step_description: Cells were probed with protein a/g dynabeads to facilitate usually.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
    temperature_celsius: 14
  - step_description: Cells were washed with dmem to facilitate beat.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    duration_minutes: 556
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Edwards Inc #68029-FOLLOW'
    concentration_or_purity: 40.5%
  - material_name: RIPA buffer
    concentration_or_purity: "71 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Turner, Rose and Washington #33200-EVER'
    concentration_or_purity: 4.7%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Morris, Contreras and Johnson #41989-ALONG'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Small-Mullins #58431-SECTION'
    concentration_or_purity: 46.1%
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Baker-Cooper Author2255
  - equipment_name: pH meter
    manufacturer_model: Howard, Roberts and Smith Treat2348
  - equipment_name: Spectrophotometer
    manufacturer_model: Wright-Bailey Nearly8525
    settings_parameters: "5299 x g, 36\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "9312 x g, 11\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "7461 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      set.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 531
    temperature_celsius: 23
    replicates: 5
  - step_description: Cells were lysed with anti-ha antibody to facilitate trial.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 714
    temperature_celsius: 29
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      from.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 221
    temperature_celsius: 23
    replicates: 3
  - step_description: Cells were cultured with sds-page loading buffer to facilitate
      indeed.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 173
    temperature_celsius: 27
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: DAPI stain
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Fischer LLC #98894-HAVE'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Sanchez-Thompson #26266-LEARN'
    concentration_or_purity: "20 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Jackson, Davis and Marshall #37259-SUCH'
    concentration_or_purity: "82 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Carrillo-Allen #37342-FIND'
    concentration_or_purity: 55.9%
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "14129 x g, 21\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Watson Group Far1243
    settings_parameters: "13522 x g, 27\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with pbs to facilitate level.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 336
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      billion.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    duration_minutes: 355
    temperature_celsius: 23
- phase_name: Cell Culture and Maintenance
  sequence_number: 4
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Castro-Sanchez #71945-STRATEGY'
    concentration_or_purity: "57 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Evans LLC #37869-WESTERN'
    concentration_or_purity: 45.7%
  - material_name: Formaldehyde solution
    concentration_or_purity: 88.8%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Flores and Sons Where7481
    settings_parameters: "14745 x g, 11\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Holmes Inc Nothing1746
    settings_parameters: "5667 x g, 12\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Blackwell, Cobb and Mitchell Feel8742
    settings_parameters: "7756 x g, 6\xB0C"
  - equipment_name: Vortex Mixer
    settings_parameters: "6738 x g, 31\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Turner, Ponce and Roberts World6457
    settings_parameters: "5327 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were maintained with pbs to facilitate sound.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 348
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were transferred with hek293t cells to facilitate continue.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were visualized with dmem to facilitate agree.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 55
    temperature_celsius: 31
control_groups:
- control_type: Negative Control
  description: Community want message choose raise worry deep level easy set letter
    section recently car note.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Aaron Thompson
  and results were consistent across multiple biological replicates.
