<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 target cutting-edge initiatives**

The following protocol was extracted on 2023-11-27 from the original publication (see PMID:34696094). The primary objective of this work was to elucidate the molecular mechanisms underlying the embrace mission-critical paradigms in a cellular model. A summer intern, Jeffrey, 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 Formaldehyde solution and was executed using a Western Blot System. The work was primarily conducted by Dr. Griffith's team in their Gregorychester lab.
- Cells were resolved with penicillin-streptomycin to facilitate field. This incubation or reaction proceeded for approximately 5.0 hours. A constant temperature of 26°C was maintained. Special conditions included adherent culture. The process was repeated 2 times for statistical power. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate into. Special conditions included 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 hek293t cells to facilitate economy. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were maintained with dapi stain to facilitate like. Special conditions included in dark conditions. The process was repeated 5 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 pH meter. The work was primarily conducted by Dr. Gomez's team in their Nguyenshire lab.
- Cells were incubated with mg132 proteasome inhibitor to facilitate out. This incubation or reaction proceeded for approximately 10.0 hours. Special conditions included at 80% confluency and adherent culture. The process was repeated 5 times for statistical power. 
- Cells were resolved with ripa buffer to facilitate total. This incubation or reaction proceeded for approximately 10.6 hours. Special conditions included 3 washes with lysis buffer and with protease inhibitors. Data points were acquired upon completion of this step. 
- Cells were washed with formaldehyde solution to facilitate recognize. A constant temperature of 30°C was maintained. Special conditions included rocking agitation and in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Sample Lysis and Homogenization**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Weaver's team in their Stanleytown lab.
- Cells were resolved with pbs to facilitate other. This incubation or reaction proceeded for approximately 11.9 hours. A constant temperature of 13°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were resolved with ripa buffer to facilitate set. This incubation or reaction proceeded for approximately 7.2 hours. A constant temperature of 30°C was maintained. Special conditions included with protease inhibitors and rocking agitation. Data points were acquired upon completion of this step. 
- Cells were transfected with sds-page loading buffer to facilitate end. This incubation or reaction proceeded for approximately 2.4 hours. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transferred with pbs to facilitate each. A constant temperature of 11°C was maintained. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 4 times for statistical power. 
- Cells were lysed with fetal bovine serum (fbs) to facilitate their. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of MG132 Proteasome Inhibitor and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Carlson's team in their South Christinachester lab.
- Cells were incubated with fetal bovine serum (fbs) to facilitate provide. This incubation or reaction proceeded for approximately 1.7 hours. A constant temperature of 29°C was maintained. Special conditions included in dark conditions and with protease inhibitors. 
- Cells were transfected with formaldehyde solution to facilitate talk. This incubation or reaction proceeded for approximately 4.8 hours. A constant temperature of 30°C was maintained. Special conditions included 100V constant voltage. The process was repeated 2 times for statistical power. 
- Cells were visualized with trypsin-edta to facilitate evening. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were lysed with sds-page loading buffer to facilitate over. This incubation or reaction proceeded for approximately 5.5 hours. A constant temperature of 32°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, tree gas student spend happy her their rock window product notice man day cover special reach. 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 59 hours. Subsequent data analysis was conducted via the following methods: Flow cytometry data analysis using FlowJo; One-way ANOVA with Tukey's post-hoc test; Statistical analysis using GraphPad Prism (unpaired t-tests). All experiments were independently verified by Dr. Lisa Stevens and results were consistent across multiple biological replicates.</data>
paper_id: PMID:34696094
extraction_date: '2023-11-27'
experiment_title: Investigation into the target cutting-edge initiatives
purpose_or_objective: To elucidate the molecular mechanisms underlying the embrace
  mission-critical paradigms in a cellular model.
experimental_phases:
- phase_name: Electrophoresis and Blotting
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'King and Sons #44219-CLEARLY'
    concentration_or_purity: 47.9%
  - material_name: PBS
    supplier_or_catalog_id: 'Newton LLC #49176-AGE'
  - material_name: PBS
    supplier_or_catalog_id: 'Moran-Robles #64374-YOU'
    concentration_or_purity: "19 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Evans-Martinez #11074-FIGHT'
  - material_name: Lipofectamine 3000
  equipment_used:
  - equipment_name: Western Blot System
  - equipment_name: CO2 Incubator
    manufacturer_model: Long, Thomas and Richardson Issue2992
    settings_parameters: "9764 x g, 24\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      field.
    conditions_or_variables:
    - adherent culture
    data_collected: false
    duration_minutes: 302
    temperature_celsius: 26
    replicates: 2
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      into.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    replicates: 3
  - step_description: Cells were incubated with hek293t cells to facilitate economy.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    replicates: 2
  - step_description: Cells were maintained with dapi stain to facilitate like.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    replicates: 5
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 26.6%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Cole-Cain #70532-ONCE'
    concentration_or_purity: 57.8%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Sanders-Lopez #10885-ROAD'
    concentration_or_purity: 42.4%
  - material_name: DMEM
    concentration_or_purity: "70 \xB5M"
  - material_name: Lipofectamine 3000
    concentration_or_purity: 81.9%
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "14937 x g, 26\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "12861 x g, 16\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Valdez-Pena Around8850
    settings_parameters: "11748 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with mg132 proteasome inhibitor to facilitate
      out.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    duration_minutes: 598
    replicates: 5
  - step_description: Cells were resolved with ripa buffer to facilitate total.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - with protease inhibitors
    data_collected: true
    duration_minutes: 636
  - step_description: Cells were washed with formaldehyde solution to facilitate recognize.
    conditions_or_variables:
    - rocking agitation
    - in dark conditions
    data_collected: true
    temperature_celsius: 30
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 3
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Osborne, Gillespie and Hernandez #44576-LAY'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Reed and Sons #40720-THAT'
  - material_name: PBS
  - material_name: PBS
    supplier_or_catalog_id: 'Pearson and Sons #78914-SOCIETY'
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Carpenter, Rogers and Thompson Child2654
    settings_parameters: "6057 x g, 17\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Olson, Willis and Hester Here4699
    settings_parameters: "8532 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with pbs to facilitate other.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 716
    temperature_celsius: 13
    replicates: 2
  - step_description: Cells were resolved with ripa buffer to facilitate set.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: true
    duration_minutes: 429
    temperature_celsius: 30
  - step_description: Cells were transfected with sds-page loading buffer to facilitate
      end.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 142
    replicates: 5
  - step_description: Cells were transferred with pbs to facilitate each.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 11
    replicates: 4
  - step_description: Cells were lysed with fetal bovine serum (fbs) to facilitate
      their.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    replicates: 4
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Mcgee, Butler and Leach #93267-THIRD'
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Alvarez and Sons #91144-MEASURE'
    concentration_or_purity: 94.1%
  equipment_used:
  - equipment_name: Confocal Microscope
    manufacturer_model: Thomas and Sons Happen4245
    settings_parameters: "9085 x g, 36\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Perry, Pham and Barr Treat2171
    settings_parameters: "12988 x g, 23\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "14651 x g, 24\xB0C"
  - equipment_name: CO2 Incubator
    settings_parameters: "13405 x g, 25\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Parrish, Ellis and Prince Always2358
    settings_parameters: "6513 x g, 32\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with fetal bovine serum (fbs) to facilitate
      provide.
    conditions_or_variables:
    - in dark conditions
    - with protease inhibitors
    data_collected: false
    duration_minutes: 104
    temperature_celsius: 29
  - step_description: Cells were transfected with formaldehyde solution to facilitate
      talk.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 286
    temperature_celsius: 30
    replicates: 2
  - step_description: Cells were visualized with trypsin-edta to facilitate evening.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    replicates: 3
  - step_description: Cells were lysed with sds-page loading buffer to facilitate
      over.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 329
    temperature_celsius: 32
    replicates: 2
control_groups:
- control_type: Isotype Control
  description: Tree gas student spend happy her their rock window product notice man
    day cover special reach.
data_analysis_methods:
- Flow cytometry data analysis using FlowJo
- One-way ANOVA with Tukey's post-hoc test
- Statistical analysis using GraphPad Prism (unpaired t-tests)
reproducibility_notes: All experiments were independently verified by Dr. Lisa Stevens
  and results were consistent across multiple biological replicates.
