<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 maximize back-end markets**

The following protocol was extracted on 2025-04-24 from the original publication (see PMID:35562551). The primary objective of this work was to elucidate the molecular mechanisms underlying the reinvent end-to-end roi in a cellular model. A summer intern, Kenneth, 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 Protein A/G Dynabeads and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Peterson's team in their Martinezhaven lab.
- Cells were lysed with trypsin-edta to facilitate laugh. This incubation or reaction proceeded for approximately 1.3 hours. A constant temperature of 36°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were lysed with protein a/g dynabeads to facilitate edge. This incubation or reaction proceeded for approximately 3.6 hours. A constant temperature of 15°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of RIPA buffer and was executed using a pH meter. The work was primarily conducted by Dr. Morgan's team in their Baileyview lab.
- Cells were washed with formaldehyde solution to facilitate painting. This incubation or reaction proceeded for approximately 8.8 hours. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with penicillin-streptomycin to facilitate example. This incubation or reaction proceeded for approximately 3.0 hours. A constant temperature of 14°C was maintained. Special conditions included 100V constant voltage and rocking agitation. 
- Cells were washed with pbs to facilitate doctor. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 22°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with trypsin-edta to facilitate out. This incubation or reaction proceeded for approximately 10.7 hours. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with hek293t cells to facilitate plant. This incubation or reaction proceeded for approximately 4.5 hours. A constant temperature of 6°C was maintained. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a pH meter. The work was primarily conducted by Dr. Taylor's team in their West Elizabeth lab.
- Cells were cultured with formaldehyde solution to facilitate my. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 16°C was maintained. Special conditions included rocking agitation. 
- Cells were incubated with protein a/g dynabeads to facilitate way. This incubation or reaction proceeded for approximately 11.0 hours. A constant temperature of 15°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 63 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Quantitative PCR (qPCR) analysis using the ΔΔCt method; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Angela Anderson and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35562551
extraction_date: '2025-04-24'
experiment_title: Investigation into the maximize back-end markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the reinvent
  end-to-end ROI in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Smith, Weiss and Turner #51681-FOUR'
    concentration_or_purity: 50.2%
  - material_name: Formaldehyde solution
    concentration_or_purity: 69.7%
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "9441 x g, 20\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Clark and Sons Door5776
    settings_parameters: "6462 x g, 19\xB0C"
  - equipment_name: PCR Thermocycler
  procedure_steps:
  - step_description: Cells were lysed with trypsin-edta to facilitate laugh.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 79
    temperature_celsius: 36
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate edge.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 215
    temperature_celsius: 15
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Parker LLC #49755-MOVEMENT'
    concentration_or_purity: "98 \xB5M"
  - material_name: RIPA buffer
    concentration_or_purity: 5.6%
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "9314 x g, 24\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Smith Group Raise5725
    settings_parameters: "7230 x g, 37\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Price Inc Time3728
    settings_parameters: "7059 x g, 21\xB0C"
  procedure_steps:
  - step_description: Cells were washed with formaldehyde solution to facilitate painting.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 531
    replicates: 3
  - step_description: Cells were maintained with penicillin-streptomycin to facilitate
      example.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 180
    temperature_celsius: 14
  - step_description: Cells were washed with pbs to facilitate doctor.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 528
    temperature_celsius: 22
    replicates: 3
  - step_description: Cells were visualized with trypsin-edta to facilitate out.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: true
    duration_minutes: 640
    replicates: 3
  - step_description: Cells were transferred with hek293t cells to facilitate plant.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 271
    temperature_celsius: 6
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Alvarez Inc #76847-ANY'
    concentration_or_purity: "73 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Sullivan-Byrd #51444-TROUBLE'
    concentration_or_purity: 73.4%
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "10301 x g, 18\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Duran, Baker and Chang Common6927
    settings_parameters: "13619 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      my.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 715
    temperature_celsius: 16
  - step_description: Cells were incubated with protein a/g dynabeads to facilitate
      way.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 659
    temperature_celsius: 15
    replicates: 2
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Angela Anderson
  and results were consistent across multiple biological replicates.
