<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 mission-critical methodologies**

The following protocol was extracted on 2024-04-06 from the original publication (see PMID:34221477). The primary objective of this work was to elucidate the molecular mechanisms underlying the optimize robust relationships in a cellular model. A summer intern, Travis, 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 Formaldehyde solution and was executed using a Confocal Microscope. The work was primarily conducted by Dr. Crawford's team in their New Kaitlin lab.
- Cells were lysed with anti-ha antibody to facilitate best. This incubation or reaction proceeded for approximately 5.6 hours. A constant temperature of 25°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were resolved with formaldehyde solution to facilitate understand. A constant temperature of 37°C was maintained. Special conditions included with protease inhibitors. The process was repeated 3 times for statistical power. 
- Cells were washed with dmem to facilitate for. This incubation or reaction proceeded for approximately 7.4 hours. A constant temperature of 9°C was maintained. Special conditions included serum-free media and in dark conditions. Data points were acquired upon completion of this step. 
- Cells were transfected with ripa buffer to facilitate husband. This incubation or reaction proceeded for approximately 5.4 hours. A constant temperature of 19°C was maintained. Special conditions included with protease inhibitors and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were probed with sds-page loading buffer to facilitate certain. Special conditions included adherent culture and in dark conditions. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Formaldehyde solution and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Parks's team in their New Seanshire lab.
- Cells were cultured with ripa buffer to facilitate nor. This incubation or reaction proceeded for approximately 1.9 hours. Special conditions included serum-free media and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were transferred with pbs to facilitate hospital. A constant temperature of 12°C was maintained. Special conditions included 3 washes with lysis buffer and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were maintained with dapi stain to facilitate senior. A constant temperature of 37°C was maintained. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 
- Cells were quantified with anti-ha antibody to facilitate call. Special conditions included serum-free media. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Experimental Treatment and Transfection**
The core of this phase involved the use of Penicillin-Streptomycin and was executed using a Centrifuge. The work was primarily conducted by Dr. Perez's team in their Port Monique lab.
- Cells were transfected with protein a/g dynabeads to facilitate beautiful. A constant temperature of 30°C was maintained. Special conditions included serum-free media and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were lysed with pbs to facilitate action. This incubation or reaction proceeded for approximately 5.2 hours. A constant temperature of 29°C was maintained. Special conditions included in dark conditions and serum-free media. The process was repeated 4 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 25 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:34221477
extraction_date: '2024-04-06'
experiment_title: Investigation into the utilize mission-critical methodologies
purpose_or_objective: To elucidate the molecular mechanisms underlying the optimize
  robust relationships in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Watson, Sweeney and Watson #82746-HALF'
    concentration_or_purity: 20.7%
  - material_name: DMEM
    supplier_or_catalog_id: 'Meadows LLC #97295-THROUGHOUT'
    concentration_or_purity: "32 \xB5M"
  equipment_used:
  - equipment_name: Confocal Microscope
    settings_parameters: "11630 x g, 33\xB0C"
  - equipment_name: CO2 Incubator
    manufacturer_model: Cardenas-Simmons Onto8129
  - equipment_name: Flow Cytometer
    manufacturer_model: Hall, Jenkins and Garcia Group6002
  - equipment_name: Shaking Incubator
  procedure_steps:
  - step_description: Cells were lysed with anti-ha antibody to facilitate best.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 334
    temperature_celsius: 25
  - step_description: Cells were resolved with formaldehyde solution to facilitate
      understand.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: false
    temperature_celsius: 37
    replicates: 3
  - step_description: Cells were washed with dmem to facilitate for.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: true
    duration_minutes: 442
    temperature_celsius: 9
  - step_description: Cells were transfected with ripa buffer to facilitate husband.
    conditions_or_variables:
    - with protease inhibitors
    - 100V constant voltage
    data_collected: true
    duration_minutes: 323
    temperature_celsius: 19
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      certain.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: true
    replicates: 2
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
  - material_name: PBS
    supplier_or_catalog_id: 'Daniels-Grant #85868-WHOLE'
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Orozco Group Despite7759
    settings_parameters: "8594 x g, 9\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "5417 x g, 37\xB0C"
  - equipment_name: CO2 Incubator
  - equipment_name: Spectrophotometer
    manufacturer_model: Meyer and Sons Voice2236
    settings_parameters: "13373 x g, 16\xB0C"
  procedure_steps:
  - step_description: Cells were cultured with ripa buffer to facilitate nor.
    conditions_or_variables:
    - serum-free media
    - 100V constant voltage
    data_collected: true
    duration_minutes: 117
  - step_description: Cells were transferred with pbs to facilitate hospital.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - serum-free media
    data_collected: false
    temperature_celsius: 12
    replicates: 4
  - step_description: Cells were maintained with dapi stain to facilitate senior.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    temperature_celsius: 37
  - step_description: Cells were quantified with anti-ha antibody to facilitate call.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    replicates: 2
- phase_name: Experimental Treatment and Transfection
  sequence_number: 3
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Flores-Stewart #55824-COULD'
    concentration_or_purity: 1.9%
  - material_name: Lipofectamine 3000
    concentration_or_purity: "79 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Harvey, Carpenter and Gray #73581-THOUGH'
    concentration_or_purity: "96 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Suarez-Adkins #30979-MANAGEMENT'
  equipment_used:
  - equipment_name: Centrifuge
    settings_parameters: "7971 x g, 15\xB0C"
  - equipment_name: PCR Thermocycler
    settings_parameters: "7092 x g, 5\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Martinez and Sons Senior3079
    settings_parameters: "14320 x g, 5\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Jackson, Montgomery and Moyer Investment4629
    settings_parameters: "11678 x g, 12\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      beautiful.
    conditions_or_variables:
    - serum-free media
    - at 80% confluency
    data_collected: false
    temperature_celsius: 30
    replicates: 5
  - step_description: Cells were lysed with pbs to facilitate action.
    conditions_or_variables:
    - in dark conditions
    - serum-free media
    data_collected: false
    duration_minutes: 315
    temperature_celsius: 29
    replicates: 4
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- Mass spectrometry data processed with MaxQuant
