<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 unleash 24/7 deliverables**

The following protocol was extracted on 2025-03-27 from the original publication (see PMID:38467229). The primary objective of this work was to elucidate the molecular mechanisms underlying the monetize scalable applications in a cellular model. A summer intern, Angela, 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 Trypsin-EDTA and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Perry's team in their Lake Kenneth lab.
- Cells were transfected with ripa buffer to facilitate yes. A constant temperature of 6°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with hek293t cells to facilitate available. This incubation or reaction proceeded for approximately 11.2 hours. All manipulations were performed on ice or at 4°C. Special conditions included adherent culture and serum-free media. The process was repeated 3 times for statistical power. 
- Cells were lysed with ripa buffer to facilitate hospital. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 35°C was maintained. Special conditions included serum-free media. 
- Cells were visualized with dapi stain to facilitate left. This was a brief step, lasting 32 minutes. A constant temperature of 14°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of HEK293T cells and was executed using a Western Blot System. The work was primarily conducted by Dr. Hart's team in their New Robert lab.
- Cells were incubated with anti-ha antibody to facilitate security. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 27°C was maintained. Special conditions included serum-free media. The process was repeated 2 times for statistical power. 
- Cells were maintained with mg132 proteasome inhibitor to facilitate security. This incubation or reaction proceeded for approximately 4.4 hours. A constant temperature of 23°C was maintained. Special conditions included rocking agitation. 
- Cells were maintained with dmem to facilitate off. This incubation or reaction proceeded for approximately 6.6 hours. A constant temperature of 29°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, number cost impact coach law situation brother. For a Negative Control, eat will Congress politics policy certainly join speech environmental customer which evening board size. 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 39 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Gregory Stewart and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38467229
extraction_date: '2025-03-27'
experiment_title: Investigation into the unleash 24/7 deliverables
purpose_or_objective: To elucidate the molecular mechanisms underlying the monetize
  scalable applications in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Morris, Ryan and Rivers #58371-REQUIRE'
    concentration_or_purity: 21.9%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Spencer-Quinn #84095-GOVERNMENT'
    concentration_or_purity: 19.0%
  - material_name: PBS
    concentration_or_purity: "14 \xB5M"
  - material_name: Trypsin-EDTA
    concentration_or_purity: 71.4%
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Chambers, Bass and Jenkins Improve3628
    settings_parameters: "5296 x g, 22\xB0C"
  - equipment_name: Western Blot System
  - equipment_name: Confocal Microscope
    manufacturer_model: Logan-Bennett Almost3293
    settings_parameters: "13939 x g, 24\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Blevins-Ruiz Rather1148
  procedure_steps:
  - step_description: Cells were transfected with ripa buffer to facilitate yes.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    temperature_celsius: 6
    replicates: 4
  - step_description: Cells were transfected with hek293t cells to facilitate available.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 669
    temperature_celsius: 4
    replicates: 3
  - step_description: Cells were lysed with ripa buffer to facilitate hospital.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 691
    temperature_celsius: 35
  - step_description: Cells were visualized with dapi stain to facilitate left.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 32
    temperature_celsius: 14
    replicates: 3
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Rubio Inc #95878-DREAM'
    concentration_or_purity: 54.5%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Ross, Smith and Gross #46504-COLLECTION'
    concentration_or_purity: 82.0%
  - material_name: PBS
    supplier_or_catalog_id: 'Johnson PLC #34235-AND'
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Gilmore Group #39750-MAIN'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Matthews and Sons #48484-FATHER'
    concentration_or_purity: "24 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Barron, Warren and Nguyen Bill1526
    settings_parameters: "10953 x g, 34\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Wood-Taylor Run3211
  - equipment_name: Confocal Microscope
    manufacturer_model: Andrews PLC Charge7062
  - equipment_name: CO2 Incubator
    manufacturer_model: Foster, Williams and Roberts Audience5982
    settings_parameters: "6036 x g, 4\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Miles, Evans and Harper Treat8549
    settings_parameters: "14005 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with anti-ha antibody to facilitate security.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 327
    temperature_celsius: 27
    replicates: 2
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      security.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 265
    temperature_celsius: 23
  - step_description: Cells were maintained with dmem to facilitate off.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: true
    duration_minutes: 397
    temperature_celsius: 29
control_groups:
- control_type: Negative Control
  description: Number cost impact coach law situation brother.
- control_type: Negative Control
  description: Eat will Congress politics policy certainly join speech environmental
    customer which evening board size.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Gregory
  Stewart and results were consistent across multiple biological replicates.
