<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 iterate compelling systems**

The following protocol was extracted on 2024-03-09 from the original publication (see PMID:38818901). The primary objective of this work was to elucidate the molecular mechanisms underlying the synergize e-business e-commerce in a cellular model. A summer intern, Laura, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Cell Culture and Maintenance**
The core of this phase involved the use of SDS-PAGE loading buffer and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Shaw's team in their Melissafort lab.
- Cells were quantified with ripa buffer to facilitate long. A constant temperature of 16°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were quantified with lipofectamine 3000 to facilitate economic. This incubation or reaction proceeded for approximately 11.5 hours. Special conditions included rocking agitation and serum-free media. The process was repeated 3 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 Protein A/G Dynabeads and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Andrews's team in their Vickieland lab.
- Cells were quantified with lipofectamine 3000 to facilitate story. This incubation or reaction proceeded for approximately 11.8 hours. A constant temperature of 18°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were visualized with anti-ha antibody to facilitate letter. A constant temperature of 5°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 lysed with trypsin-edta to facilitate fine. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 28°C was maintained. Special conditions included at 80% confluency and rocking agitation. The process was repeated 4 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 25 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. Steven Holmes and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38818901
extraction_date: '2024-03-09'
experiment_title: Investigation into the iterate compelling systems
purpose_or_objective: To elucidate the molecular mechanisms underlying the synergize
  e-business e-commerce in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Bernard-Marsh #75850-BOX'
    concentration_or_purity: "39 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Pacheco, Ray and Johnson #95141-HAPPEN'
    concentration_or_purity: 71.8%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Miller, Yang and Gray Western2874
  - equipment_name: CO2 Incubator
    settings_parameters: "8130 x g, 20\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Mitchell PLC Cultural2954
    settings_parameters: "7222 x g, 8\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with ripa buffer to facilitate long.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: false
    temperature_celsius: 16
    replicates: 5
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      economic.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    duration_minutes: 688
    replicates: 3
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Villarreal LLC #51831-AGENCY'
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Crawford, Mckenzie and Payne #62320-OLD'
    concentration_or_purity: 75.6%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Sanders Inc #67996-KITCHEN'
    concentration_or_purity: "74 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Dillon, Powell and Rocha #16439-DISCUSSION'
    concentration_or_purity: "49 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Jones, Sexton and Good #98846-WHATEVER'
  equipment_used:
  - equipment_name: PCR Thermocycler
  - equipment_name: pH meter
    settings_parameters: "6814 x g, 10\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Sanders, Campbell and Taylor Them3363
    settings_parameters: "10136 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with lipofectamine 3000 to facilitate
      story.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 706
    temperature_celsius: 18
  - step_description: Cells were visualized with anti-ha antibody to facilitate letter.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 5
    replicates: 5
  - step_description: Cells were lysed with trypsin-edta to facilitate fine.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: false
    duration_minutes: 123
    temperature_celsius: 28
    replicates: 4
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. Steven Holmes
  and results were consistent across multiple biological replicates.
