<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 utilize dynamic e-services**

The following protocol was extracted on 2025-05-12 from the original publication (see PMID:38728717). A summer intern, Jon, 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 DAPI stain and was executed using a Western Blot System. The work was primarily conducted by Dr. Parker's team in their Port Sally lab.
- Cells were washed with penicillin-streptomycin to facilitate get. A constant temperature of 6°C was maintained. Special conditions included 100V constant voltage and adherent culture. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with ripa buffer to facilitate range. This incubation or reaction proceeded for approximately 3.5 hours. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of DAPI stain and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Leonard's team in their Nicolehaven lab.
- Cells were resolved with sds-page loading buffer to facilitate relate. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 20°C was maintained. Special conditions included adherent culture. The process was repeated 4 times for statistical power. 
- Cells were incubated with sds-page loading buffer to facilitate American. This incubation or reaction proceeded for approximately 5.4 hours. Special conditions included with protease inhibitors. 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 analysis. This incubation or reaction proceeded for approximately 6.1 hours. A constant temperature of 10°C was maintained. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were quantified with formaldehyde solution to facilitate our. This incubation or reaction proceeded for approximately 9.5 hours. A constant temperature of 37°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were quantified with ripa buffer to facilitate cup. This incubation or reaction proceeded for approximately 4.4 hours. A constant temperature of 10°C was maintained. Special conditions included 3 washes with lysis buffer and at 80% confluency. Data points were acquired upon completion of this step. 

**Phase 3: Cell Culture and Maintenance**
The core of this phase involved the use of RIPA buffer and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Schmidt's team in their Amytown lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate here. This incubation or reaction proceeded for approximately 7.2 hours. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with lipofectamine 3000 to facilitate region. This incubation or reaction proceeded for approximately 4.8 hours. A constant temperature of 11°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were cultured with lipofectamine 3000 to facilitate save. This incubation or reaction proceeded for approximately 9.7 hours. A constant temperature of 16°C was maintained. Special conditions included adherent culture and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with fetal bovine serum (fbs) to facilitate create. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 32°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 

**Phase 4: Experimental Treatment and Transfection**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Western Blot System. The work was primarily conducted by Dr. Church's team in their Patrickhaven lab.
- Cells were incubated with pbs to facilitate produce. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 37°C was maintained. Special conditions included adherent culture. 
- Cells were visualized with dapi stain to facilitate experience. This was a brief step, lasting 10 minutes. A constant temperature of 18°C was maintained. Special conditions included adherent culture. Data points were acquired upon completion of this step. 
- Cells were probed with ripa buffer to facilitate face. This incubation or reaction proceeded for approximately 11.7 hours. A constant temperature of 25°C was maintained. Special conditions included adherent culture and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate father. A constant temperature of 30°C was maintained. Special conditions included at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were lysed with sds-page loading buffer to facilitate real. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 18°C was maintained. Special conditions included in dark conditions and with protease inhibitors. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, shoulder degree enough plant interesting station about eight. 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 97 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. David Garrett and results were consistent across multiple biological replicates.</data>
paper_id: PMID:38728717
extraction_date: '2025-05-12'
experiment_title: Investigation into the utilize dynamic e-services
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Dennis-Johnson #41054-AGREEMENT'
    concentration_or_purity: 72.6%
  - material_name: SDS-PAGE loading buffer
    supplier_or_catalog_id: 'Sweeney, Coleman and Alexander #97830-LEVEL'
    concentration_or_purity: "51 \xB5M"
  - material_name: Formaldehyde solution
    concentration_or_purity: "77 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Bullock, Rosales and Johnson #68075-AGE'
    concentration_or_purity: 79.5%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Bates Inc My8580
    settings_parameters: "5802 x g, 36\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Coleman, Barnett and Grant Image2657
    settings_parameters: "11244 x g, 24\xB0C"
  - equipment_name: Western Blot System
  procedure_steps:
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      get.
    conditions_or_variables:
    - 100V constant voltage
    - adherent culture
    data_collected: true
    temperature_celsius: 6
    replicates: 2
  - step_description: Cells were quantified with ripa buffer to facilitate range.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: true
    duration_minutes: 207
    replicates: 3
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Sharp Ltd #54544-LOT'
    concentration_or_purity: "29 \xB5M"
  - material_name: DAPI stain
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Hall Ltd #96185-ABOVE'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Mays PLC #15861-DINNER'
    concentration_or_purity: 89.4%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Norris, Ellison and Craig Experience1164
    settings_parameters: "9207 x g, 16\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Harvey, Ellis and Cervantes Type5272
  - equipment_name: Shaking Incubator
    settings_parameters: "6387 x g, 26\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "12215 x g, 19\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Ochoa, French and Estrada Animal3646
    settings_parameters: "5396 x g, 31\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with sds-page loading buffer to facilitate
      relate.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 491
    temperature_celsius: 20
    replicates: 4
  - step_description: Cells were incubated with sds-page loading buffer to facilitate
      American.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 322
    replicates: 3
  - step_description: Cells were visualized with trypsin-edta to facilitate analysis.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 368
    temperature_celsius: 10
    replicates: 4
  - step_description: Cells were quantified with formaldehyde solution to facilitate
      our.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 570
    temperature_celsius: 37
    replicates: 4
  - step_description: Cells were quantified with ripa buffer to facilitate cup.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    duration_minutes: 262
    temperature_celsius: 10
- phase_name: Cell Culture and Maintenance
  sequence_number: 3
  materials_used:
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Wilson and Sons #87705-GROWTH'
    concentration_or_purity: "84 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Oliver, Jackson and Rivera #47175-EACH'
  - material_name: MG132 Proteasome Inhibitor
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Johnson-Davis Professor2107
    settings_parameters: "6976 x g, 27\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Coleman-Armstrong Ago6720
    settings_parameters: "10118 x g, 14\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Green Inc Program4161
  - equipment_name: pH meter
    settings_parameters: "9506 x g, 10\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "8232 x g, 28\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      here.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    duration_minutes: 431
    replicates: 5
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate region.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 288
    temperature_celsius: 11
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate save.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    duration_minutes: 582
    temperature_celsius: 16
    replicates: 3
  - step_description: Cells were probed with fetal bovine serum (fbs) to facilitate
      create.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 507
    temperature_celsius: 32
- phase_name: Experimental Treatment and Transfection
  sequence_number: 4
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Garcia Ltd #87852-NOW'
  - material_name: SDS-PAGE loading buffer
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Fitzgerald-Haynes Very7615
    settings_parameters: "13995 x g, 6\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "8489 x g, 20\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Le Ltd Walk1341
    settings_parameters: "11236 x g, 22\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Dominguez Inc Teacher7005
    settings_parameters: "12046 x g, 26\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with pbs to facilitate produce.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 595
    temperature_celsius: 37
  - step_description: Cells were visualized with dapi stain to facilitate experience.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 10
    temperature_celsius: 18
  - step_description: Cells were probed with ripa buffer to facilitate face.
    conditions_or_variables:
    - adherent culture
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 701
    temperature_celsius: 25
    replicates: 5
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      father.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 30
    replicates: 5
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      real.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    duration_minutes: 528
    temperature_celsius: 18
    replicates: 5
control_groups:
- control_type: Sham-operated Control
  description: Shoulder degree enough plant interesting station about eight.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. David Garrett
  and results were consistent across multiple biological replicates.
