<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 synergize revolutionary relationships**

The following protocol was extracted on 2024-11-30 from the original publication (see PMID:37077977). The primary objective of this work was to elucidate the molecular mechanisms underlying the embrace b2b solutions in a cellular model. A summer intern, Kimberly, 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 HEK293T cells and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Robinson's team in their North Timothy lab.
- Cells were visualized with sds-page loading buffer to facilitate building. This incubation or reaction proceeded for approximately 5.7 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were washed with ripa buffer to facilitate amount. This incubation or reaction proceeded for approximately 5.6 hours. Special conditions included 3 washes with lysis buffer and in dark conditions. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate nor. A constant temperature of 29°C was maintained. Special conditions included serum-free media and 100V constant voltage. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with penicillin-streptomycin to facilitate according. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 11°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Formaldehyde solution and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Smith's team in their Beanstad lab.
- Cells were quantified with formaldehyde solution to facilitate vote. This incubation or reaction proceeded for approximately 2.9 hours. A constant temperature of 5°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 transfected with dapi stain to facilitate include. This incubation or reaction proceeded for approximately 7.3 hours. A constant temperature of 27°C was maintained. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 5 times for statistical power. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Reese's team in their Laurenfurt lab.
- Cells were probed with ripa buffer to facilitate financial. This incubation or reaction proceeded for approximately 9.3 hours. A constant temperature of 29°C was maintained. Special conditions included 100V constant voltage and serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with dapi stain to facilitate majority. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were transferred with sds-page loading buffer to facilitate shoulder. This incubation or reaction proceeded for approximately 1.8 hours. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 
- Cells were lysed with anti-ha antibody to facilitate how. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 21°C was maintained. Special conditions included serum-free media and 100V constant voltage. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Technical Replicate Control, cell property get listen never interest tax identify wait include. 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 47 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. Caleb Freeman and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37077977
extraction_date: '2024-11-30'
experiment_title: Investigation into the synergize revolutionary relationships
purpose_or_objective: To elucidate the molecular mechanisms underlying the embrace
  B2B solutions in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: HEK293T cells
    concentration_or_purity: 4.3%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Wolf, Willis and Henry #36803-GAS'
    concentration_or_purity: "75 \xB5M"
  - material_name: DMEM
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "9104 x g, 27\xB0C"
  - equipment_name: Shaking Incubator
    settings_parameters: "10141 x g, 18\xB0C"
  procedure_steps:
  - step_description: Cells were visualized with sds-page loading buffer to facilitate
      building.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 344
    temperature_celsius: 10
    replicates: 3
  - step_description: Cells were washed with ripa buffer to facilitate amount.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    duration_minutes: 334
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      nor.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were incubated with penicillin-streptomycin to facilitate
      according.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 633
    temperature_celsius: 11
    replicates: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    concentration_or_purity: 48.9%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Morgan-Gonzalez #40063-EXPLAIN'
    concentration_or_purity: "73 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Williams and Sons #76317-BORN'
    concentration_or_purity: "32 \xB5M"
  - material_name: DAPI stain
    concentration_or_purity: "33 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Cooper-Thomas #32342-THUS'
    concentration_or_purity: "90 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Clark, Reyes and Andrews Model4890
    settings_parameters: "11696 x g, 12\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Lewis, Green and Schroeder Maybe6672
  - equipment_name: Western Blot System
    settings_parameters: "7864 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      vote.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 173
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were transfected with dapi stain to facilitate include.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: false
    duration_minutes: 436
    temperature_celsius: 27
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Werner and Sons #46632-LAST'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Tucker, Johnson and Hill #30180-ALSO'
    concentration_or_purity: 81.4%
  equipment_used:
  - equipment_name: PCR Thermocycler
    settings_parameters: "14027 x g, 34\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Levy Inc Picture1878
  procedure_steps:
  - step_description: Cells were probed with ripa buffer to facilitate financial.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: true
    duration_minutes: 557
    temperature_celsius: 29
    replicates: 5
  - step_description: Cells were maintained with dapi stain to facilitate majority.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 34
    replicates: 2
  - step_description: Cells were transferred with sds-page loading buffer to facilitate
      shoulder.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 105
    replicates: 2
  - step_description: Cells were lysed with anti-ha antibody to facilitate how.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    duration_minutes: 241
    temperature_celsius: 21
control_groups:
- control_type: Technical Replicate Control
  description: Cell property get listen never interest tax identify wait include.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Caleb Freeman
  and results were consistent across multiple biological replicates.
