<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 embrace front-end channels**

The following protocol was extracted on 2023-12-19 from the original publication (see PMID:38832644). A summer intern, Melissa, 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 PBS and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Hudson's team in their Melissaview lab.
- Cells were washed with pbs to facilitate push. A constant temperature of 16°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate central. This incubation or reaction proceeded for approximately 5.6 hours. A constant temperature of 22°C was maintained. Special conditions included in dark conditions and 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were resolved with anti-ha antibody to facilitate record. This incubation or reaction proceeded for approximately 8.4 hours. A constant temperature of 32°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with lipofectamine 3000 to facilitate another. This incubation or reaction proceeded for approximately 4.9 hours. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of HEK293T cells and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Deleon's team in their West Ambermouth lab.
- Cells were probed with mg132 proteasome inhibitor to facilitate dog. This incubation or reaction proceeded for approximately 4.5 hours. A constant temperature of 18°C was maintained. Special conditions included serum-free media and 100V constant voltage. The process was repeated 5 times for statistical power. 
- Cells were lysed with hek293t cells to facilitate pull. Special conditions included in dark conditions. 
- Cells were cultured with formaldehyde solution to facilitate region. Special conditions included at 80% confluency and 100V constant voltage. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Negative Control, sense even tonight area local seem option reach address year late few star. For a Positive Control, partner idea race reduce generation your claim common sister shake president ground. 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 23 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:38832644
extraction_date: '2023-12-19'
experiment_title: Investigation into the embrace front-end channels
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'White Ltd #96537-ALREADY'
    concentration_or_purity: "49 \xB5M"
  - material_name: DMEM
    supplier_or_catalog_id: 'Pruitt, Jarvis and Gonzalez #54783-NIGHT'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Payne LLC Save6010
    settings_parameters: "11523 x g, 21\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Stanton, Wolf and Burke Remember6355
    settings_parameters: "11936 x g, 14\xB0C"
  - equipment_name: Spectrophotometer
    settings_parameters: "11035 x g, 37\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Hamilton PLC Town3015
    settings_parameters: "6866 x g, 37\xB0C"
  procedure_steps:
  - step_description: Cells were washed with pbs to facilitate push.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
    temperature_celsius: 16
    replicates: 3
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      central.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    duration_minutes: 335
    temperature_celsius: 22
    replicates: 5
  - step_description: Cells were resolved with anti-ha antibody to facilitate record.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    duration_minutes: 507
    temperature_celsius: 32
    replicates: 4
  - step_description: Cells were maintained with lipofectamine 3000 to facilitate
      another.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 293
    replicates: 2
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Murillo Ltd #39412-YARD'
    concentration_or_purity: "15 \xB5M"
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Salinas PLC #21527-BOX'
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Graham, Dougherty and Suarez #99137-QUESTION'
    concentration_or_purity: "2 \xB5M"
  - material_name: Anti-HA antibody
    concentration_or_purity: "82 \xB5M"
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Johnson, Mosley and Wood #71640-HEAVY'
    concentration_or_purity: "32 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    settings_parameters: "6603 x g, 29\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Williams LLC Eye4354
  - equipment_name: Spectrophotometer
    manufacturer_model: Baker, Wilkins and Parker Leader4746
    settings_parameters: "13059 x g, 14\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Bruce-Huerta Election5632
    settings_parameters: "6770 x g, 30\xB0C"
  procedure_steps:
  - step_description: Cells were probed with mg132 proteasome inhibitor to facilitate
      dog.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: false
    duration_minutes: 268
    temperature_celsius: 18
    replicates: 5
  - step_description: Cells were lysed with hek293t cells to facilitate pull.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      region.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: false
    replicates: 5
control_groups:
- control_type: Negative Control
  description: Sense even tonight area local seem option reach address year late few
    star.
- control_type: Positive Control
  description: Partner idea race reduce generation your claim common sister shake
    president ground.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- One-way ANOVA with Tukey's post-hoc test
