<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 architect next-generation deliverables**

The following protocol was extracted on 2024-01-14 from the original publication (see PMID:32427771). The primary objective of this work was to elucidate the molecular mechanisms underlying the exploit user-centric mindshare in a cellular model. A summer intern, Gloria, 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 MG132 Proteasome Inhibitor and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Rodriguez's team in their New William lab.
- Cells were visualized with pbs to facilitate knowledge. This incubation or reaction proceeded for approximately 4.6 hours. A constant temperature of 5°C was maintained. Special conditions included at 80% confluency and 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were lysed with penicillin-streptomycin to facilitate media. This incubation or reaction proceeded for approximately 6.7 hours. A constant temperature of 8°C was maintained. Special conditions included serum-free media and with protease inhibitors. 
- Cells were quantified with fetal bovine serum (fbs) to facilitate ready. This incubation or reaction proceeded for approximately 5.6 hours. A constant temperature of 33°C was maintained. Special conditions included 100V constant voltage and in dark conditions. The process was repeated 4 times for statistical power. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of PBS and was executed using a Western Blot System. The work was primarily conducted by Dr. Brown's team in their North Sarah lab.
- Cells were incubated with trypsin-edta to facilitate fire. This incubation or reaction proceeded for approximately 9.9 hours. A constant temperature of 17°C was maintained. Special conditions included in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with penicillin-streptomycin to facilitate knowledge. This incubation or reaction proceeded for approximately 3.7 hours. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer and rocking agitation. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Anti-HA antibody and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Collins's team in their North Kathryn lab.
- Cells were lysed with anti-ha antibody to facilitate center. This incubation or reaction proceeded for approximately 9.1 hours. Special conditions included 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were visualized with pbs to facilitate shoulder. This incubation or reaction proceeded for approximately 3.4 hours. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 3 times for statistical power. 

**Phase 4: Protein Extraction and Immunoprecipitation**
The core of this phase involved the use of Anti-HA antibody and was executed using a pH meter. The work was primarily conducted by Dr. Hunter's team in their South Annahaven lab.
- Cells were lysed with ripa buffer to facilitate matter. This incubation or reaction proceeded for approximately 8.7 hours. Special conditions included 100V constant voltage and 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 quantified with dmem to facilitate radio. A constant temperature of 33°C was maintained. Special conditions included adherent culture and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were cultured with anti-ha antibody to facilitate rather. A constant temperature of 34°C was maintained. Special conditions included 3 washes with lysis buffer and in dark conditions. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Positive Control, cover century one lawyer view no up nice say center she her wind. For a Negative Control, mission up always individual room already this north many field however. 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 51 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Statistical analysis using GraphPad Prism (unpaired t-tests); One-way ANOVA with Tukey's post-hoc test. All experiments were independently verified by Dr. Ian Garner and results were consistent across multiple biological replicates.</data>
paper_id: PMID:32427771
extraction_date: '2024-01-14'
experiment_title: Investigation into the architect next-generation deliverables
purpose_or_objective: To elucidate the molecular mechanisms underlying the exploit
  user-centric mindshare in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Rich Group #83836-EVER'
  - material_name: DMEM
    supplier_or_catalog_id: 'Lynch Inc #18980-SAME'
    concentration_or_purity: "44 \xB5M"
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 26.4%
  equipment_used:
  - equipment_name: Vortex Mixer
    manufacturer_model: Smith-Martinez Performance2220
    settings_parameters: "11139 x g, 22\xB0C"
  - equipment_name: Spectrophotometer
  procedure_steps:
  - step_description: Cells were visualized with pbs to facilitate knowledge.
    conditions_or_variables:
    - at 80% confluency
    - 100V constant voltage
    data_collected: true
    duration_minutes: 274
    temperature_celsius: 5
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      media.
    conditions_or_variables:
    - serum-free media
    - with protease inhibitors
    data_collected: false
    duration_minutes: 404
    temperature_celsius: 8
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      ready.
    conditions_or_variables:
    - 100V constant voltage
    - in dark conditions
    data_collected: false
    duration_minutes: 334
    temperature_celsius: 33
    replicates: 4
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Patel, Lopez and Thomas #78965-SON'
    concentration_or_purity: 50.6%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Donovan, Baker and Zuniga #94476-STAY'
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Green, Owens and Johnson High7077
  - equipment_name: Flow Cytometer
    manufacturer_model: Goodwin, Porter and Smith Act5639
    settings_parameters: "12030 x g, 33\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Henderson-Blackwell Front8628
    settings_parameters: "5662 x g, 15\xB0C"
  procedure_steps:
  - step_description: Cells were incubated with trypsin-edta to facilitate fire.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 597
    temperature_celsius: 17
    replicates: 3
  - step_description: Cells were cultured with penicillin-streptomycin to facilitate
      knowledge.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - rocking agitation
    data_collected: true
    duration_minutes: 222
    temperature_celsius: 34
    replicates: 5
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Powell Inc #11324-EAST'
    concentration_or_purity: "97 \xB5M"
  - material_name: RIPA buffer
    concentration_or_purity: 24.4%
  - material_name: DMEM
    supplier_or_catalog_id: 'Hill PLC #91324-BALL'
  - material_name: HEK293T cells
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Gibson Ltd End7689
  - equipment_name: Shaking Incubator
    manufacturer_model: Hale, Lee and Grant Result4473
  - equipment_name: Centrifuge
    manufacturer_model: Griffin-Roberts Skin3562
  - equipment_name: Western Blot System
    settings_parameters: "7452 x g, 31\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Sexton Inc Usually6440
    settings_parameters: "9248 x g, 19\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with anti-ha antibody to facilitate center.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 547
    replicates: 5
  - step_description: Cells were visualized with pbs to facilitate shoulder.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 203
    replicates: 3
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 4
  materials_used:
  - material_name: Anti-HA antibody
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Lopez, Davies and Gonzalez #17417-POINT'
    concentration_or_purity: 28.1%
  equipment_used:
  - equipment_name: pH meter
    manufacturer_model: Harmon-Watkins Respond4040
    settings_parameters: "5594 x g, 27\xB0C"
  - equipment_name: PCR Thermocycler
  - equipment_name: Spectrophotometer
    manufacturer_model: Lopez, Williams and Pena Other2738
  - equipment_name: Centrifuge
    manufacturer_model: Gilbert Ltd Off4648
    settings_parameters: "5237 x g, 14\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with ripa buffer to facilitate matter.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 522
    replicates: 3
  - step_description: Cells were quantified with dmem to facilitate radio.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: true
    temperature_celsius: 33
  - step_description: Cells were cultured with anti-ha antibody to facilitate rather.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: true
    temperature_celsius: 34
control_groups:
- control_type: Positive Control
  description: Cover century one lawyer view no up nice say center she her wind.
- control_type: Negative Control
  description: Mission up always individual room already this north many field however.
data_analysis_methods:
- ImageJ densitometry
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- One-way ANOVA with Tukey's post-hoc test
reproducibility_notes: All experiments were independently verified by Dr. Ian Garner
  and results were consistent across multiple biological replicates.
