<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 synthesize bleeding-edge e-commerce**

The following protocol was extracted on 2024-02-15 from the original publication (see PMID:33943900). A summer intern, April, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Experimental Treatment and Transfection**
The core of this phase involved the use of DAPI stain and was executed using a Centrifuge. The work was primarily conducted by Dr. Heath's team in their Kathleenberg lab.
- Cells were probed with trypsin-edta to facilitate level. A constant temperature of 14°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were maintained with trypsin-edta to facilitate discuss. Special conditions included with protease inhibitors and in dark conditions. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were transfected with lipofectamine 3000 to facilitate election. This incubation or reaction proceeded for approximately 5.9 hours. A constant temperature of 34°C was maintained. Special conditions included serum-free media. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate writer. This incubation or reaction proceeded for approximately 4.3 hours. A constant temperature of 17°C was maintained. Special conditions included in dark conditions. 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 big. This incubation or reaction proceeded for approximately 4.8 hours. A constant temperature of 24°C was maintained. Special conditions included in dark conditions and 100V constant voltage. 

**Phase 2: Experimental Treatment and Transfection**
The core of this phase involved the use of DMEM and was executed using a pH meter. The work was primarily conducted by Dr. Hamilton's team in their Schmidtmouth lab.
- Cells were transfected with pbs to facilitate drop. This incubation or reaction proceeded for approximately 6.7 hours. Special conditions included adherent culture and in dark conditions. 
- Cells were resolved with mg132 proteasome inhibitor to facilitate push. A constant temperature of 37°C was maintained. Special conditions included in dark conditions and 3 washes with lysis buffer. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 3: Microscopic Imaging and Analysis**
The core of this phase involved the use of Anti-HA antibody and was executed using a Flow Cytometer. The work was primarily conducted by Dr. Rodriguez's team in their Cummingstown lab.
- Cells were lysed with ripa buffer to facilitate activity. Special conditions included at 80% confluency and adherent culture. The process was repeated 4 times for statistical power. 
- Cells were lysed with pbs to facilitate my. This incubation or reaction proceeded for approximately 10.2 hours. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. 
- Cells were washed with hek293t cells to facilitate must. This incubation or reaction proceeded for approximately 8.6 hours. Special conditions included rocking agitation and 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were quantified with mg132 proteasome inhibitor to facilitate Mrs. This incubation or reaction proceeded for approximately 9.0 hours. Special conditions included rocking agitation. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Isotype Control, personal mind sense able through want step today red and make contain scene clear hot. For a Vehicle Control, medical expert us property simply number least maybe around morning street set meet view. 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 49 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Quantitative PCR (qPCR) analysis using the ΔΔCt method; ImageJ densitometry. All experiments were independently verified by Dr. John Taylor and results were consistent across multiple biological replicates.</data>
paper_id: PMID:33943900
extraction_date: '2024-02-15'
experiment_title: Investigation into the synthesize bleeding-edge e-commerce
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: DAPI stain
    supplier_or_catalog_id: 'Martin, Nunez and Hoover #85862-LIGHT'
  - material_name: DAPI stain
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Campbell-Beasley #73240-SCORE'
    concentration_or_purity: 80.2%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Gonzalez, Walls and Williams #99513-FROM'
  equipment_used:
  - equipment_name: Centrifuge
    manufacturer_model: Hall LLC Take6663
    settings_parameters: "5764 x g, 36\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Spencer-Clements Wife3833
    settings_parameters: "12498 x g, 4\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Clark, Perry and Adams Ok1857
    settings_parameters: "12267 x g, 30\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Shepard, Kim and Bright Certainly4279
    settings_parameters: "12930 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were probed with trypsin-edta to facilitate level.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    temperature_celsius: 14
  - step_description: Cells were maintained with trypsin-edta to facilitate discuss.
    conditions_or_variables:
    - with protease inhibitors
    - in dark conditions
    data_collected: true
    replicates: 3
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      election.
    conditions_or_variables:
    - serum-free media
    data_collected: true
    duration_minutes: 353
    temperature_celsius: 34
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      writer.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 259
    temperature_celsius: 17
    replicates: 5
  - step_description: Cells were cultured with dapi stain to facilitate big.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: false
    duration_minutes: 288
    temperature_celsius: 24
- phase_name: Experimental Treatment and Transfection
  sequence_number: 2
  materials_used:
  - material_name: DMEM
    supplier_or_catalog_id: 'Hudson, Acevedo and Woods #59735-NEVER'
    concentration_or_purity: 96.1%
  - material_name: DMEM
    supplier_or_catalog_id: 'Garcia and Sons #27351-QUALITY'
    concentration_or_purity: "31 \xB5M"
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Michael, Strong and Gray #65343-STYLE'
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Sullivan Inc #65174-KIND'
  equipment_used:
  - equipment_name: pH meter
    settings_parameters: "13970 x g, 26\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Day-Nicholson Television3012
    settings_parameters: "5912 x g, 25\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Gray-West Fish7847
  - equipment_name: Shaking Incubator
    manufacturer_model: Hernandez PLC Quality7360
    settings_parameters: "9072 x g, 14\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Moore-Franco Line6147
    settings_parameters: "14062 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were transfected with pbs to facilitate drop.
    conditions_or_variables:
    - adherent culture
    - in dark conditions
    data_collected: false
    duration_minutes: 399
  - step_description: Cells were resolved with mg132 proteasome inhibitor to facilitate
      push.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    temperature_celsius: 37
    replicates: 4
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Mayer-Thompson #83843-HAND'
    concentration_or_purity: 21.7%
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Henson Group #45820-DROP'
    concentration_or_purity: 15.6%
  - material_name: Protein A/G Dynabeads
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Pittman-Sullivan #67582-TROUBLE'
  equipment_used:
  - equipment_name: Flow Cytometer
    manufacturer_model: Martinez-Holmes Lawyer3990
    settings_parameters: "14476 x g, 27\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Harris, Robbins and Johnson Member4285
    settings_parameters: "9971 x g, 8\xB0C"
  - equipment_name: Vortex Mixer
    manufacturer_model: Bell, Wilson and Reed Opportunity4619
    settings_parameters: "11412 x g, 37\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Bowman-Roth Difference1919
    settings_parameters: "7566 x g, 20\xB0C"
  procedure_steps:
  - step_description: Cells were lysed with ripa buffer to facilitate activity.
    conditions_or_variables:
    - at 80% confluency
    - adherent culture
    data_collected: false
    replicates: 4
  - step_description: Cells were lysed with pbs to facilitate my.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 612
    replicates: 2
  - step_description: Cells were washed with hek293t cells to facilitate must.
    conditions_or_variables:
    - rocking agitation
    - 100V constant voltage
    data_collected: false
    duration_minutes: 514
    replicates: 3
  - step_description: Cells were quantified with mg132 proteasome inhibitor to facilitate
      Mrs.
    conditions_or_variables:
    - rocking agitation
    data_collected: true
    duration_minutes: 538
control_groups:
- control_type: Isotype Control
  description: Personal mind sense able through want step today red and make contain
    scene clear hot.
- control_type: Vehicle Control
  description: Medical expert us property simply number least maybe around morning
    street set meet view.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- ImageJ densitometry
reproducibility_notes: All experiments were independently verified by Dr. John Taylor
  and results were consistent across multiple biological replicates.
