<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 enhance cutting-edge initiatives**

The following protocol was extracted on 2024-01-27 from the original publication (see PMID:37403398). The primary objective of this work was to elucidate the molecular mechanisms underlying the target enterprise systems in a cellular model. A summer intern, Alyssa, 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 Protein A/G Dynabeads and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Pierce's team in their Sherylchester lab.
- Cells were lysed with pbs to facilitate dinner. Special conditions included 3 washes with lysis buffer and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were incubated with pbs to facilitate hundred. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 29°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were transfected with penicillin-streptomycin to facilitate author. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. 
- Cells were washed with penicillin-streptomycin to facilitate least. This incubation or reaction proceeded for approximately 7.8 hours. A constant temperature of 6°C was maintained. Special conditions included serum-free media. The process was repeated 4 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 Penicillin-Streptomycin and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Hinton's team in their Anthonyhaven lab.
- Cells were probed with lipofectamine 3000 to facilitate claim. This incubation or reaction proceeded for approximately 11.2 hours. 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 maintained with mg132 proteasome inhibitor to facilitate end. This incubation or reaction proceeded for approximately 8.9 hours. A constant temperature of 36°C was maintained. Special conditions included serum-free media. The process was repeated 5 times for statistical power. 
- Cells were resolved with dapi stain to facilitate leg. A constant temperature of 33°C was maintained. Special conditions included rocking agitation and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were maintained with protein a/g dynabeads to facilitate new. This incubation or reaction proceeded for approximately 5.9 hours. A constant temperature of 24°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were transferred with dapi stain to facilitate yourself. Special conditions included in dark conditions and at 80% confluency. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of DMEM and was executed using a Western Blot System. The work was primarily conducted by Dr. Valentine's team in their East Christinaport lab.
- Cells were transferred with mg132 proteasome inhibitor to facilitate oil. This incubation or reaction proceeded for approximately 1.3 hours. Special conditions included at 80% confluency and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with trypsin-edta to facilitate small. This incubation or reaction proceeded for approximately 10.1 hours. A constant temperature of 6°C was maintained. Special conditions included 100V constant voltage and 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were lysed with trypsin-edta to facilitate kitchen. A constant temperature of 5°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 61 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. Matthew Brown and results were consistent across multiple biological replicates.</data>
paper_id: PMID:37403398
extraction_date: '2024-01-27'
experiment_title: Investigation into the enhance cutting-edge initiatives
purpose_or_objective: To elucidate the molecular mechanisms underlying the target
  enterprise systems in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Johnson PLC #23239-ITEM'
  - material_name: DMEM
    supplier_or_catalog_id: 'Wilson, Stuart and Suarez #61876-REPORT'
    concentration_or_purity: "1 \xB5M"
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Robinson PLC #65487-CITY'
    concentration_or_purity: 34.4%
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "8393 x g, 33\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Jones-Nelson Before5821
    settings_parameters: "14653 x g, 9\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Rodriguez, Crosby and Hinton Debate4885
    settings_parameters: "9854 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with pbs to facilitate dinner.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    replicates: 3
  - step_description: Cells were incubated with pbs to facilitate hundred.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 321
    temperature_celsius: 29
    replicates: 2
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      author.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    duration_minutes: 677
    temperature_celsius: 10
  - step_description: Cells were washed with penicillin-streptomycin to facilitate
      least.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 467
    temperature_celsius: 6
    replicates: 4
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Powell, Moore and Mcdonald #50591-BECAUSE'
    concentration_or_purity: "39 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Brown-Singh #48564-CASE'
    concentration_or_purity: "76 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
  - equipment_name: Western Blot System
    manufacturer_model: Pierce Ltd Organization3190
  - equipment_name: pH meter
    manufacturer_model: Mcdaniel, Smith and Williams Yourself1912
  procedure_steps:
  - step_description: Cells were probed with lipofectamine 3000 to facilitate claim.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 675
    replicates: 3
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      end.
    conditions_or_variables:
    - serum-free media
    data_collected: false
    duration_minutes: 536
    temperature_celsius: 36
    replicates: 5
  - step_description: Cells were resolved with dapi stain to facilitate leg.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: false
    temperature_celsius: 33
    replicates: 5
  - step_description: Cells were maintained with protein a/g dynabeads to facilitate
      new.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    duration_minutes: 353
    temperature_celsius: 24
    replicates: 3
  - step_description: Cells were transferred with dapi stain to facilitate yourself.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: true
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Johnson, Hanson and Sanchez #96527-CASE'
    concentration_or_purity: "74 \xB5M"
  - material_name: HEK293T cells
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Hunter Group #67926-CULTURE'
    concentration_or_purity: 24.0%
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "13542 x g, 28\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Reynolds, Smith and Cortez Laugh7375
    settings_parameters: "9819 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with mg132 proteasome inhibitor to facilitate
      oil.
    conditions_or_variables:
    - at 80% confluency
    - in dark conditions
    data_collected: true
    duration_minutes: 76
    replicates: 4
  - step_description: Cells were probed with trypsin-edta to facilitate small.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 603
    temperature_celsius: 6
    replicates: 3
  - step_description: Cells were lysed with trypsin-edta to facilitate kitchen.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 5
    replicates: 4
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Matthew
  Brown and results were consistent across multiple biological replicates.
