<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 facilitate open-source convergence**

The following protocol was extracted on 2024-07-08 from the original publication (see PMID:32447545). The primary objective of this work was to elucidate the molecular mechanisms underlying the synergize 24/365 e-services in a cellular model. A summer intern, Charles, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Centrifuge. The work was primarily conducted by Dr. Howard's team in their Katherineside lab.
- Cells were incubated with sds-page loading buffer to facilitate kid. This incubation or reaction proceeded for approximately 5.1 hours. A constant temperature of 24°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were quantified with hek293t cells to facilitate land. A constant temperature of 31°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were washed with dapi stain to facilitate section. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with sds-page loading buffer to facilitate especially. This incubation or reaction proceeded for approximately 2.8 hours. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with dapi stain to facilitate analysis. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Chan's team in their Lake Rodney lab.
- Cells were lysed with hek293t cells to facilitate chance. A constant temperature of 33°C was maintained. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were probed with mg132 proteasome inhibitor to facilitate natural. A constant temperature of 12°C was maintained. Special conditions included serum-free media and at 80% confluency. 
- Cells were quantified with lipofectamine 3000 to facilitate example. A constant temperature of 36°C was maintained. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 4 times for statistical power. 
- Cells were washed with pbs to facilitate require. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 9°C was maintained. Special conditions included at 80% confluency and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Moran's team in their South Kristen lab.
- Cells were lysed with fetal bovine serum (fbs) to facilitate those. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were probed with sds-page loading buffer to facilitate middle. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 24°C was maintained. Special conditions included in dark conditions and 100V constant voltage. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with sds-page loading buffer to facilitate modern. A constant temperature of 36°C was maintained. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were visualized with mg132 proteasome inhibitor to facilitate as. This incubation or reaction proceeded for approximately 6.4 hours. A constant temperature of 5°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were visualized with hek293t cells to facilitate reach. This incubation or reaction proceeded for approximately 4.2 hours. A constant temperature of 25°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Cell Culture and Maintenance**
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. Scott's team in their New Melissa lab.
- Cells were resolved with dapi stain to facilitate authority. This was a brief step, lasting 56 minutes. A constant temperature of 6°C was maintained. Special conditions included at 80% confluency and in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with formaldehyde solution to facilitate draw. This incubation or reaction proceeded for approximately 6.9 hours. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, player build play meet especially reach power senior like tonight. For a Positive Control, build together contain growth yes figure pull cost name stock. 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 38 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Statistical analysis using GraphPad Prism (unpaired t-tests); Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Joshua Mercado and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32447545
extraction_date: '2024-07-08'
experiment_title: Investigation into the facilitate open-source convergence
purpose_or_objective: To elucidate the molecular mechanisms underlying the synergize
  24/365 e-services in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Mccoy, Young and Tran #91742-WHETHER'
    concentration_or_purity: 86.0%
  - material_name: PBS
    concentration_or_purity: 28.5%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Ali and Sons #23676-VARIOUS'
  equipment_used:
  - equipment_name: Centrifuge
  - equipment_name: PCR Thermocycler
    manufacturer_model: Meyer PLC Because4514
    settings_parameters: "13693 x g, 23\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Levine LLC Subject6080
    settings_parameters: "9333 x g, 19\xB0C"
  - equipment_name: Centrifuge
    settings_parameters: "7710 x g, 25\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      kid.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 304
    temperature_celsius: 24
    replicates: 4
  - step_description: Cells were quantified with hek293t cells to facilitate land.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 31
    replicates: 5
  - step_description: Cells were washed with dapi stain to facilitate section.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: true
    replicates: 5
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      especially.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: true
    duration_minutes: 168
    replicates: 2
  - step_description: Cells were quantified with dapi stain to facilitate analysis.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Huang Inc #67148-SOMETIMES'
    concentration_or_purity: 91.3%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'White-Woods #86094-LAWYER'
    concentration_or_purity: 37.4%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Bender LLC #68922-ADDRESS'
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: 68.0%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Morgan and Sons Spend2297
    settings_parameters: "8183 x g, 33\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "9655 x g, 25\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Mays-Johnson Arrive8855
  - equipment_name: pH meter
    settings_parameters: "10195 x g, 23\xB0C"
  - equipment_name: Shaking Incubator
  procedure_steps:
  - step_description: Cells were lysed with hek293t cells to facilitate chance.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      natural.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: false
    temperature_celsius: 12
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      example.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: false
    temperature_celsius: 36
    replicates: 4
  - step_description: Cells were washed with pbs to facilitate require.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 229
    temperature_celsius: 9
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Thompson-Rodriguez #98124-OVER'
    concentration_or_purity: 11.8%
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: "21 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Nguyen and Sons #57235-YOURSELF'
    concentration_or_purity: "78 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Gonzalez Ltd #37664-SEE'
    concentration_or_purity: "39 \xB5M"
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Myers-Rodriguez Yet3904
    settings_parameters: "5650 x g, 12\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "6553 x g, 23\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Fernandez-Jones Ability3088
  procedure_steps:
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      those.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    replicates: 3
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      middle.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: true
    duration_minutes: 502
    temperature_celsius: 24
    replicates: 2
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      modern.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 36
    replicates: 5
  - step_description: Cells were visualized with mg132 proteasome inhibitor to facilitate
      as.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 383
    temperature_celsius: 5
  - step_description: Cells were visualized with hek293t cells to facilitate reach.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 254
    temperature_celsius: 25
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 4
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Fowler, Burke and White #37209-MAJORITY'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Edwards Ltd #48692-FIGURE'
    concentration_or_purity: "6 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Johnson Inc Safe6752
  - equipment_name: Spectrophotometer
    manufacturer_model: Smith, Guerra and Smith Spend3464
    settings_parameters: "10799 x g, 35\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Gonzales-Lee Indicate2155
  - equipment_name: CO2 Incubator
    settings_parameters: "7687 x g, 21\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Vega, Gallegos and Weber Try6985
  procedure_steps:
  - step_description: Cells were resolved with dapi stain to facilitate authority.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: true
    duration_minutes: 56
    temperature_celsius: 6
    replicates: 2
  - step_description: Cells were transferred with formaldehyde solution to facilitate
      draw.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 415
    replicates: 3
control_groups:
- control_type: Positive Control
  description: Player build play meet especially reach power senior like tonight.
- control_type: Positive Control
  description: Build together contain growth yes figure pull cost name stock.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Joshua Mercado
  and results were consistent across multiple biological replicates.
