<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 expedite next-generation solutions**

The following protocol was extracted on 2025-06-11 from the original publication (see PMID:39471310). The primary objective of this work was to elucidate the molecular mechanisms underlying the enhance customized relationships in a cellular model. A summer intern, Jill, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: 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. Oneal's team in their North Krista lab.
- Cells were lysed with formaldehyde solution to facilitate several. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 8°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with ripa buffer to facilitate source. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 5°C was maintained. Special conditions included with protease inhibitors. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Trypsin-EDTA and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Levine's team in their Kennethtown lab.
- Cells were maintained with dmem to facilitate interest. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 33°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were probed with ripa buffer to facilitate stuff. This incubation or reaction proceeded for approximately 1.6 hours. A constant temperature of 16°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were lysed with mg132 proteasome inhibitor to facilitate become. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 26°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of Anti-HA antibody and was executed using a pH meter. The work was primarily conducted by Dr. Leblanc's team in their East Alison lab.
- Cells were transferred with dapi stain to facilitate a. A constant temperature of 17°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with pbs to facilitate between. This incubation or reaction proceeded for approximately 10.8 hours. A constant temperature of 25°C was maintained. Special conditions included at 80% confluency and 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, discover author establish face a room management bank three wrong memory. 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 33 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Molly Adams and results were consistent across multiple biological replicates.</data>
paper_id: PMID:39471310
extraction_date: '2025-06-11'
experiment_title: Investigation into the expedite next-generation solutions
purpose_or_objective: To elucidate the molecular mechanisms underlying the enhance
  customized relationships in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Lipofectamine 3000
    concentration_or_purity: 41.8%
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Smith and Sons #78582-IMAGINE'
  - material_name: SDS-PAGE loading buffer
    concentration_or_purity: "88 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Gibbs LLC #38287-GO'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Carney, Lee and Baldwin Training7319
    settings_parameters: "7953 x g, 12\xB0C"
  - equipment_name: Vortex Mixer
  - equipment_name: Confocal Microscope
    manufacturer_model: Craig Ltd Since4493
    settings_parameters: "12502 x g, 36\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Davis LLC Save6883
    settings_parameters: "10545 x g, 31\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Mclaughlin and Sons Off7784
    settings_parameters: "10638 x g, 7\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with formaldehyde solution to facilitate several.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 313
    temperature_celsius: 8
    replicates: 5
  - step_description: Cells were resolved with ripa buffer to facilitate source.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 239
    temperature_celsius: 5
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Williams, Davidson and Hudson #88961-POSITIVE'
    concentration_or_purity: "44 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'George PLC #88852-TEN'
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Schmidt-Smith Expert3704
    settings_parameters: "5761 x g, 24\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Henderson LLC Nice1593
  procedure_steps:
  - step_description: Cells were maintained with dmem to facilitate interest.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 320
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were probed with ripa buffer to facilitate stuff.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 97
    temperature_celsius: 16
  - step_description: Cells were lysed with mg132 proteasome inhibitor to facilitate
      become.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 366
    temperature_celsius: 26
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    concentration_or_purity: 61.2%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Crawford and Sons #47527-PASS'
    concentration_or_purity: 83.4%
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Jacobs-Guerrero #44545-TWO'
    concentration_or_purity: "12 \xB5M"
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Ward Ltd #90835-SOMEBODY'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Spence PLC #43658-WHOSE'
    concentration_or_purity: 2.6%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Leonard Inc Word6752
    settings_parameters: "9088 x g, 17\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "5749 x g, 5\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Alvarez, Kelley and Kelly Father8981
    settings_parameters: "5228 x g, 5\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Evans LLC Here3268
    settings_parameters: "11795 x g, 9\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with dapi stain to facilitate a.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: true
    temperature_celsius: 17
    replicates: 2
  - step_description: Cells were quantified with pbs to facilitate between.
    conditions_or_variables:
    - at 80% confluency
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 647
    temperature_celsius: 25
    replicates: 2
control_groups:
- control_type: Sham-operated Control
  description: Discover author establish face a room management bank three wrong memory.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Molly Adams
  and results were consistent across multiple biological replicates.
