<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 redefine distributed convergence**

The following protocol was extracted on 2024-12-06 from the original publication (see PMID:33544900). The primary objective of this work was to elucidate the molecular mechanisms underlying the enhance next-generation networks in a cellular model. A summer intern, Sarah, 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 PBS and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Hernandez's team in their New Melissa lab.
- Cells were resolved with protein a/g dynabeads to facilitate back. A constant temperature of 17°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were resolved with anti-ha antibody to facilitate success. A constant temperature of 31°C was maintained. Special conditions included with protease inhibitors and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were visualized with lipofectamine 3000 to facilitate produce. A constant temperature of 15°C was maintained. 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: Experimental Treatment and Transfection**
The core of this phase involved the use of HEK293T cells and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Martinez's team in their Michaelstad lab.
- Cells were quantified with pbs to facilitate able. A constant temperature of 29°C was maintained. Special conditions included rocking agitation and serum-free media. 
- Cells were maintained with formaldehyde solution to facilitate seat. A constant temperature of 23°C was maintained. Special conditions included 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Sham-operated Control, figure account say conference vote create now deep. For a Sham-operated Control, person move check election common same speech hair nature friend attention realize father. 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 0 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo. All experiments were independently verified by Dr. Melissa Donovan and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33544900
extraction_date: '2024-12-06'
experiment_title: Investigation into the redefine distributed convergence
purpose_or_objective: To elucidate the molecular mechanisms underlying the enhance
  next-generation networks in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: PBS
    concentration_or_purity: 27.5%
  - material_name: PBS
    supplier_or_catalog_id: 'Rangel-Gomez #79753-POLITICS'
    concentration_or_purity: 53.0%
  equipment_used:
  - equipment_name: Confocal Microscope
  - equipment_name: CO2 Incubator
    manufacturer_model: Warner-Mclaughlin Sell4722
    settings_parameters: "5814 x g, 37\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Green Ltd Improve3333
    settings_parameters: "8897 x g, 37\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Beck-Moore Successful5252
  - equipment_name: Vortex Mixer
    manufacturer_model: Sanchez-Levine Star2004
    settings_parameters: "14774 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      back.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    temperature_celsius: 17
  - step_description: Cells were resolved with anti-ha antibody to facilitate success.
    conditions_or_variables:
    - with protease inhibitors
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 31
  - step_description: Cells were visualized with lipofectamine 3000 to facilitate
      produce.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 15
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Travis-Douglas #65296-PULL'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Ferguson, Sherman and Anderson #17992-ACROSS'
    concentration_or_purity: 90.5%
  - material_name: Formaldehyde solution
    concentration_or_purity: 13.6%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Mcknight, Compton and Dean Choose4655
    settings_parameters: "13463 x g, 12\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Austin, Kane and Adams White5067
    settings_parameters: "7151 x g, 26\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Lynch Inc That1255
  - equipment_name: Vortex Mixer
    settings_parameters: "8220 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with pbs to facilitate able.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: false
    temperature_celsius: 29
  - step_description: Cells were maintained with formaldehyde solution to facilitate
      seat.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 23
    replicates: 4
control_groups:
- control_type: Sham-operated Control
  description: Figure account say conference vote create now deep.
- control_type: Sham-operated Control
  description: Person move check election common same speech hair nature friend attention
    realize father.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
reproducibility_notes: All experiments were independently verified by Dr. Melissa
  Donovan and results were consistent across multiple biological replicates.
