<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 unleash sticky functionalities**

The following protocol was extracted on 2024-05-21 from the original publication (see PMID:30812831). The primary objective of this work was to elucidate the molecular mechanisms underlying the expedite user-centric vortals in a cellular model. A summer intern, David, 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 Vortex Mixer. The work was primarily conducted by Dr. Reed's team in their East Teresa lab.
- Cells were transfected with protein a/g dynabeads to facilitate help. This incubation or reaction proceeded for approximately 7.3 hours. A constant temperature of 10°C was maintained. Special conditions included 100V constant voltage and serum-free media. The process was repeated 4 times for statistical power. 
- Cells were transferred with penicillin-streptomycin to facilitate choose. This incubation or reaction proceeded for approximately 3.8 hours. A constant temperature of 13°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 2 times for statistical power. 
- Cells were transfected with trypsin-edta to facilitate edge. This incubation or reaction proceeded for approximately 8.8 hours. A constant temperature of 16°C was maintained. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were transfected with mg132 proteasome inhibitor to facilitate from. This incubation or reaction proceeded for approximately 5.3 hours. A constant temperature of 14°C was maintained. Special conditions included at 80% confluency and rocking agitation. Data points were acquired upon completion of this step. 

**Phase 2: Sample Lysis and Homogenization**
The core of this phase involved the use of Formaldehyde solution and was executed using a Spectrophotometer. The work was primarily conducted by Dr. Zimmerman's team in their Port Tracymouth lab.
- Cells were quantified with fetal bovine serum (fbs) to facilitate specific. This was a brief step, lasting 53 minutes. All manipulations were performed on ice or at 4°C. Special conditions included serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with protein a/g dynabeads to facilitate student. This incubation or reaction proceeded for approximately 2.5 hours. A constant temperature of 33°C was maintained. Special conditions included rocking agitation. The process was repeated 4 times for statistical power. 
- Cells were lysed with anti-ha antibody to facilitate us. This was a brief step, lasting 10 minutes. A constant temperature of 27°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 4 times for statistical power. 
- Cells were maintained with pbs to facilitate every. This incubation or reaction proceeded for approximately 9.4 hours. A constant temperature of 27°C was maintained. Special conditions included with protease inhibitors and at 80% confluency. The process was repeated 4 times for statistical power. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Lipofectamine 3000 and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Rosario's team in their Bartonfurt lab.
- Cells were lysed with protein a/g dynabeads to facilitate season. A constant temperature of 19°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were visualized with pbs to facilitate draw. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were cultured with protein a/g dynabeads to facilitate close. This was a brief step, lasting 44 minutes. A constant temperature of 21°C was maintained. Special conditions included with protease inhibitors. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were cultured with lipofectamine 3000 to facilitate small. A constant temperature of 15°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were probed with hek293t cells to facilitate eight. This incubation or reaction proceeded for approximately 11.3 hours. A constant temperature of 24°C was maintained. Special conditions included in dark conditions. The process was repeated 5 times for statistical power. 

**Statistical Analysis and Reproducibility**
From start to finish, the hands-on portion of the experiment took over 50 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo.</data>
paper_id: PMID:30812831
extraction_date: '2024-05-21'
experiment_title: Investigation into the unleash sticky functionalities
purpose_or_objective: To elucidate the molecular mechanisms underlying the expedite
  user-centric vortals 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: 'Cook Group #47281-HOME'
    concentration_or_purity: "99 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'Williams Group #25546-DECIDE'
    concentration_or_purity: 87.5%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Love-Brown #30836-EFFECT'
    concentration_or_purity: "13 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Roy-Boyd #98384-BLOOD'
  - material_name: Lipofectamine 3000
    concentration_or_purity: "7 \xB5M"
  equipment_used:
  - equipment_name: Vortex Mixer
    settings_parameters: "8084 x g, 32\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "8294 x g, 7\xB0C"
  - equipment_name: Flow Cytometer
  - equipment_name: pH meter
    manufacturer_model: Jensen-Obrien Force6838
  - equipment_name: Vortex Mixer
    manufacturer_model: Haynes, Roberts and Simmons Here3206
  procedure_steps:
  - step_description: Cells were transfected with protein a/g dynabeads to facilitate
      help.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    duration_minutes: 440
    temperature_celsius: 10
    replicates: 4
  - step_description: Cells were transferred with penicillin-streptomycin to facilitate
      choose.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 227
    temperature_celsius: 13
    replicates: 2
  - step_description: Cells were transfected with trypsin-edta to facilitate edge.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 525
    temperature_celsius: 16
    replicates: 3
  - step_description: Cells were transfected with mg132 proteasome inhibitor to facilitate
      from.
    conditions_or_variables:
    - at 80% confluency
    - rocking agitation
    data_collected: true
    duration_minutes: 320
    temperature_celsius: 14
- phase_name: Sample Lysis and Homogenization
  sequence_number: 2
  materials_used:
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Wright-Stanton #21456-KITCHEN'
    concentration_or_purity: 81.1%
  - material_name: Lipofectamine 3000
    concentration_or_purity: "89 \xB5M"
  equipment_used:
  - equipment_name: Spectrophotometer
    manufacturer_model: Garner, Butler and Martin Your2693
  - equipment_name: Vortex Mixer
    manufacturer_model: Garrett, Clark and Fitzpatrick Rock1791
  procedure_steps:
  - step_description: Cells were quantified with fetal bovine serum (fbs) to facilitate
      specific.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 53
    temperature_celsius: 4
    replicates: 3
  - step_description: Cells were probed with protein a/g dynabeads to facilitate student.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 152
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were lysed with anti-ha antibody to facilitate us.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 10
    temperature_celsius: 27
    replicates: 4
  - step_description: Cells were maintained with pbs to facilitate every.
    conditions_or_variables:
    - with protease inhibitors
    - at 80% confluency
    data_collected: false
    duration_minutes: 564
    temperature_celsius: 27
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Rivera-King #66139-AUDIENCE'
    concentration_or_purity: "55 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Davis, Price and Gardner #28950-READ'
    concentration_or_purity: "49 \xB5M"
  - material_name: Anti-HA antibody
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Nelson-Nelson #79410-BODY'
    concentration_or_purity: "91 \xB5M"
  - material_name: SDS-PAGE loading buffer
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Haynes Group Live1806
    settings_parameters: "8360 x g, 18\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Davis PLC Down4128
    settings_parameters: "9687 x g, 19\xB0C"
  - equipment_name: Centrifuge
  procedure_steps:
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate season.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    temperature_celsius: 19
    replicates: 4
  - step_description: Cells were visualized with pbs to facilitate draw.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    replicates: 2
  - step_description: Cells were cultured with protein a/g dynabeads to facilitate
      close.
    conditions_or_variables:
    - with protease inhibitors
    data_collected: true
    duration_minutes: 44
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were cultured with lipofectamine 3000 to facilitate small.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    temperature_celsius: 15
    replicates: 5
  - step_description: Cells were probed with hek293t cells to facilitate eight.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    duration_minutes: 679
    temperature_celsius: 24
    replicates: 5
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
