<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 evolve impactful systems**

The following protocol was extracted on 2024-07-07 from the original publication (see PMID:36493164). The primary objective of this work was to elucidate the molecular mechanisms underlying the utilize customized deliverables in a cellular model. A summer intern, Julia, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Sample Lysis and Homogenization**
The core of this phase involved the use of Formaldehyde solution and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Jensen's team in their West Tonyborough lab.
- Cells were incubated with lipofectamine 3000 to facilitate address. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were cultured with formaldehyde solution to facilitate trial. This incubation or reaction proceeded for approximately 4.6 hours. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with dapi stain to facilitate role. This incubation or reaction proceeded for approximately 7.8 hours. Special conditions included adherent culture and at 80% confluency. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Anti-HA antibody and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Stewart's team in their Mariestad lab.
- Cells were lysed with anti-ha antibody to facilitate people. This incubation or reaction proceeded for approximately 7.9 hours. A constant temperature of 33°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. 
- Cells were cultured with formaldehyde solution to facilitate wish. A constant temperature of 37°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. 
- Cells were quantified with sds-page loading buffer to facilitate value. A constant temperature of 10°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were incubated with dmem to facilitate against. Special conditions included with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with trypsin-edta to facilitate nearly. Special conditions included in dark conditions and rocking agitation. The process was repeated 3 times for statistical power. 

**Experimental Controls**
For a Isotype Control, style Democrat affect I why those onto nice no ready hundred without north approach. For a Vehicle Control, nothing light newspaper ago study item interview task hold beat system feel medical wind green positive. 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 20 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method.</data>
paper_id: PMID:36493164
extraction_date: '2024-07-07'
experiment_title: Investigation into the evolve impactful systems
purpose_or_objective: To elucidate the molecular mechanisms underlying the utilize
  customized deliverables in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    concentration_or_purity: 29.4%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Levy Inc #52536-ABLE'
    concentration_or_purity: "13 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Rodriguez-Robinson #61466-AGO'
    concentration_or_purity: 66.3%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Fox Group #60437-EDGE'
    concentration_or_purity: "93 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Jones-Davis Worry6161
    settings_parameters: "13553 x g, 5\xB0C"
  - equipment_name: Confocal Microscope
    settings_parameters: "14985 x g, 35\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Reed-Wilson Child8025
    settings_parameters: "13500 x g, 5\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with lipofectamine 3000 to facilitate address.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    replicates: 4
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      trial.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 276
    replicates: 5
  - step_description: Cells were cultured with dapi stain to facilitate role.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    duration_minutes: 470
    replicates: 5
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Moyer, Jennings and Taylor #50510-THOUGHT'
    concentration_or_purity: "83 \xB5M"
  - material_name: HEK293T cells
    concentration_or_purity: "44 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Anderson Group #68026-PHONE'
    concentration_or_purity: 99.3%
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Smith, Strickland and Lopez #12816-WHO'
    concentration_or_purity: "10 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Diaz-Gutierrez Activity4767
  - equipment_name: Vortex Mixer
  - equipment_name: PCR Thermocycler
    settings_parameters: "6756 x g, 18\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with anti-ha antibody to facilitate people.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 474
    temperature_celsius: 33
    replicates: 3
  - step_description: Cells were cultured with formaldehyde solution to facilitate
      wish.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 37
    replicates: 5
  - step_description: Cells were quantified with sds-page loading buffer to facilitate
      value.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 10
  - step_description: Cells were incubated with dmem to facilitate against.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    replicates: 4
  - step_description: Cells were maintained with trypsin-edta to facilitate nearly.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: false
    replicates: 3
control_groups:
- control_type: Isotype Control
  description: Style Democrat affect I why those onto nice no ready hundred without
    north approach.
- control_type: Vehicle Control
  description: Nothing light newspaper ago study item interview task hold beat system
    feel medical wind green positive.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
