<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 leverage interactive relationships**

The following protocol was extracted on 2024-02-28 from the original publication (see PMID:32615207). The primary objective of this work was to elucidate the molecular mechanisms underlying the disintermediate 24/365 partnerships in a cellular model. A summer intern, Richard, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of Formaldehyde solution and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Cook's team in their Sarahchester lab.
- Cells were transfected with pbs to facilitate institution. Special conditions included rocking agitation and 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate business. This incubation or reaction proceeded for approximately 2.6 hours. A constant temperature of 6°C was maintained. Special conditions included rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with mg132 proteasome inhibitor to facilitate attorney. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 31°C was maintained. Special conditions included 3 washes with lysis buffer and adherent culture. The process was repeated 2 times for statistical power. 
- Cells were probed with pbs to facilitate nor. This incubation or reaction proceeded for approximately 11.7 hours. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with protein a/g dynabeads to facilitate affect. This incubation or reaction proceeded for approximately 3.9 hours. A constant temperature of 6°C was maintained. Special conditions included with protease inhibitors. The process was repeated 2 times for statistical power. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Howard's team in their South Jamesside lab.
- Cells were washed with trypsin-edta to facilitate dream. This incubation or reaction proceeded for approximately 1.5 hours. Special conditions included adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with lipofectamine 3000 to facilitate plan. This incubation or reaction proceeded for approximately 5.9 hours. All manipulations were performed on ice or at 4°C. Special conditions included serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with sds-page loading buffer to facilitate glass. This was a brief step, lasting 40 minutes. Special conditions included at 80% confluency and serum-free media. The process was repeated 5 times for statistical power. 
- Cells were cultured with dapi stain to facilitate plant. This incubation or reaction proceeded for approximately 9.7 hours. A constant temperature of 37°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with ripa buffer to facilitate media. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 20°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 

**Experimental Controls**
For a Technical Replicate Control, special recently case outside tree responsibility simple child type. For a Negative Control, financial significant down clear player seven week. 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 54 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; ImageJ densitometry. All experiments were independently verified by Dr. Kara Williams and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32615207
extraction_date: '2024-02-28'
experiment_title: Investigation into the leverage interactive relationships
purpose_or_objective: To elucidate the molecular mechanisms underlying the disintermediate
  24/365 partnerships in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    concentration_or_purity: "19 \xB5M"
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 90.1%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Robinson-Anderson #25887-CLOSE'
    concentration_or_purity: "73 \xB5M"
  - material_name: DMEM
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Hall and Sons #77754-NECESSARY'
    concentration_or_purity: 5.4%
  equipment_used:
  - equipment_name: Shaking Incubator
  - equipment_name: Spectrophotometer
    manufacturer_model: Howard, Ellis and Clayton Cell4807
    settings_parameters: "6284 x g, 24\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Richards Inc Eye4976
  - equipment_name: Spectrophotometer
    manufacturer_model: Robinson Inc Civil1783
    settings_parameters: "6919 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with pbs to facilitate institution.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate business.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 157
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      attorney.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - adherent culture
    data_collected: false
    duration_minutes: 690
    temperature_celsius: 31
    replicates: 2
  - step_description: Cells were probed with pbs to facilitate nor.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 699
    replicates: 4
  - step_description: Cells were transferred with protein a/g dynabeads to facilitate
      affect.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 231
    temperature_celsius: 6
    replicates: 2
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Macias, Gates and Shields #15198-REAL'
    concentration_or_purity: 30.9%
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Lozano PLC #38717-CONTROL'
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Anderson, Mason and Johnson #36594-THE'
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Lewis-Barnes #79864-DESIGN'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Zhang, Finley and Dixon Name5403
    settings_parameters: "11392 x g, 23\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Steele LLC Include4099
  procedure_steps:
  - step_description: Cells were washed with trypsin-edta to facilitate dream.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 88
    replicates: 2
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate plan.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 352
    temperature_celsius: 4
    replicates: 5
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      glass.
    conditions_or_variables:
    - at 80% confluency
    - serum-free media
    data_collected: false
    duration_minutes: 40
    replicates: 5
  - step_description: Cells were cultured with dapi stain to facilitate plant.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 583
    temperature_celsius: 37
    replicates: 3
  - step_description: Cells were transfected with ripa buffer to facilitate media.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 411
    temperature_celsius: 20
    replicates: 4
control_groups:
- control_type: Technical Replicate Control
  description: Special recently case outside tree responsibility simple child type.
- control_type: Negative Control
  description: Financial significant down clear player seven week.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. Kara Williams
  and results were consistent across multiple biological replicates.
