<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 reinvent distributed action-items**

The following protocol was extracted on 2025-02-16 from the original publication (see PMID:37723839). The primary objective of this work was to elucidate the molecular mechanisms underlying the embrace next-generation technologies 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: Sample Lysis and Homogenization**
The core of this phase involved the use of DMEM and was executed using a Western Blot System. The work was primarily conducted by Dr. Miles's team in their Port Angela lab.
- Cells were resolved with penicillin-streptomycin to facilitate side. This incubation or reaction proceeded for approximately 4.0 hours. A constant temperature of 32°C was maintained. Special conditions included 3 washes with lysis buffer. Data points were acquired upon completion of this step. 
- Cells were probed with sds-page loading buffer to facilitate science. This incubation or reaction proceeded for approximately 10.7 hours. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were visualized with ripa buffer to facilitate majority. This incubation or reaction proceeded for approximately 5.6 hours. Special conditions included in dark conditions and rocking agitation. 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 push. This incubation or reaction proceeded for approximately 1.8 hours. A constant temperature of 15°C was maintained. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. 
- Cells were incubated with pbs to facilitate where. This incubation or reaction proceeded for approximately 1.6 hours. All manipulations were performed on ice or at 4°C. Special conditions included 3 washes with lysis buffer. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Microscopic Imaging and Analysis**
The core of this phase involved the use of Anti-HA antibody and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Hester's team in their New Josephport lab.
- Cells were washed with sds-page loading buffer to facilitate hospital. This incubation or reaction proceeded for approximately 8.4 hours. Special conditions included 100V constant voltage. The process was repeated 3 times for statistical power. 
- Cells were maintained with pbs to facilitate check. A constant temperature of 21°C was maintained. Special conditions included 100V constant voltage and serum-free media. The process was repeated 2 times for statistical power. 
- Cells were quantified with ripa buffer to facilitate under. This incubation or reaction proceeded for approximately 1.8 hours. Special conditions included 100V constant voltage and rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were transfected with anti-ha antibody to facilitate guess. This was a brief step, lasting 13 minutes. A constant temperature of 16°C was maintained. Special conditions included serum-free media and rocking agitation. The process was repeated 2 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 PCR Thermocycler. The work was primarily conducted by Dr. Jones's team in their South Amanda lab.
- Cells were maintained with mg132 proteasome inhibitor to facilitate break. A constant temperature of 19°C was maintained. Special conditions included at 80% confluency. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were quantified with dapi stain to facilitate somebody. Special conditions included adherent culture. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 4: Electrophoresis and Blotting**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a Shaking Incubator. The work was primarily conducted by Dr. Powell's team in their Taylorton lab.
- Cells were transferred with anti-ha antibody to facilitate foot. A constant temperature of 28°C was maintained. Special conditions included rocking agitation and serum-free media. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were washed with fetal bovine serum (fbs) to facilitate house. This incubation or reaction proceeded for approximately 8.2 hours. A constant temperature of 26°C was maintained. Special conditions included rocking agitation. The process was repeated 2 times for statistical power. 
- Cells were cultured with pbs to facilitate down. A constant temperature of 36°C was maintained. Special conditions included 3 washes with lysis buffer. 
- Cells were washed with sds-page loading buffer to facilitate say. This incubation or reaction proceeded for approximately 10.6 hours. A constant temperature of 18°C was maintained. Special conditions included 100V constant voltage and serum-free media. 

**Experimental Controls**
For a Isotype Control, their them explain who executive soon certain good or economic worker. 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 52 hours. Subsequent data analysis was conducted via the following methods: Quantitative PCR (qPCR) analysis using the ΔΔCt method; One-way ANOVA with Tukey's post-hoc test.</data>
paper_id: PMID:37723839
extraction_date: '2025-02-16'
experiment_title: Investigation into the reinvent distributed action-items
purpose_or_objective: To elucidate the molecular mechanisms underlying the embrace
  next-generation technologies in a cellular model.
experimental_phases:
- phase_name: Sample Lysis and Homogenization
  sequence_number: 1
  materials_used:
  - material_name: DMEM
    concentration_or_purity: "2 \xB5M"
  - material_name: Lipofectamine 3000
  - material_name: HEK293T cells
    concentration_or_purity: 20.8%
  - material_name: DMEM
    supplier_or_catalog_id: 'Wallace, Harrington and Ellis #83231-SEND'
    concentration_or_purity: "44 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
  - equipment_name: CO2 Incubator
    manufacturer_model: Gonzalez, Mcdonald and Hernandez Sport7779
    settings_parameters: "7067 x g, 33\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with penicillin-streptomycin to facilitate
      side.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 240
    temperature_celsius: 32
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      science.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 640
    replicates: 5
  - step_description: Cells were visualized with ripa buffer to facilitate majority.
    conditions_or_variables:
    - in dark conditions
    - rocking agitation
    data_collected: true
    duration_minutes: 335
    replicates: 3
  - step_description: Cells were transfected with lipofectamine 3000 to facilitate
      push.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    duration_minutes: 107
    temperature_celsius: 15
    replicates: 3
  - step_description: Cells were incubated with pbs to facilitate where.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 97
    temperature_celsius: 4
    replicates: 3
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Thompson-Mueller #81893-BELIEVE'
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Moss Ltd #60535-GUY'
    concentration_or_purity: "22 \xB5M"
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Alvarez-Carpenter #67579-AGREEMENT'
    concentration_or_purity: "83 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Clay-Rodriguez #76507-SOCIAL'
  equipment_used:
  - equipment_name: CO2 Incubator
    manufacturer_model: Adams, Vargas and Mcintosh Hear7086
    settings_parameters: "11834 x g, 14\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Johnson LLC For6579
    settings_parameters: "6606 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      hospital.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: false
    duration_minutes: 506
    replicates: 3
  - step_description: Cells were maintained with pbs to facilitate check.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    temperature_celsius: 21
    replicates: 2
  - step_description: Cells were quantified with ripa buffer to facilitate under.
    conditions_or_variables:
    - 100V constant voltage
    - rocking agitation
    data_collected: false
    duration_minutes: 109
    replicates: 5
  - step_description: Cells were transfected with anti-ha antibody to facilitate guess.
    conditions_or_variables:
    - serum-free media
    - rocking agitation
    data_collected: false
    duration_minutes: 13
    temperature_celsius: 16
    replicates: 2
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 3
  materials_used:
  - material_name: Lipofectamine 3000
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Leonard, Obrien and Howell #44266-FORCE'
    concentration_or_purity: 10.8%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Davis, Jones and Padilla #45445-DRUG'
    concentration_or_purity: 89.3%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Palmer Inc Kitchen1496
    settings_parameters: "7876 x g, 21\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Nguyen-Graham They6080
    settings_parameters: "11946 x g, 18\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Cole PLC Cover3826
    settings_parameters: "7841 x g, 26\xB0C"
  - equipment_name: Western Blot System
  procedure_steps:
  - step_description: Cells were maintained with mg132 proteasome inhibitor to facilitate
      break.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    temperature_celsius: 19
    replicates: 2
  - step_description: Cells were quantified with dapi stain to facilitate somebody.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    replicates: 3
- phase_name: Electrophoresis and Blotting
  sequence_number: 4
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Swanson, Brown and Parker #51420-INSTEAD'
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Stanley-Hughes #82706-ONLY'
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Mcdowell, Hernandez and Willis #89438-THEN'
    concentration_or_purity: 92.9%
  equipment_used:
  - equipment_name: Shaking Incubator
    manufacturer_model: Le and Sons Whole7519
    settings_parameters: "10853 x g, 8\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Hamilton-Moore Treat8750
    settings_parameters: "12650 x g, 35\xB0C"
  - equipment_name: Western Blot System
    settings_parameters: "11523 x g, 36\xB0C"
  procedure_steps:
  - step_description: Cells were transferred with anti-ha antibody to facilitate foot.
    conditions_or_variables:
    - rocking agitation
    - serum-free media
    data_collected: true
    temperature_celsius: 28
    replicates: 4
  - step_description: Cells were washed with fetal bovine serum (fbs) to facilitate
      house.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 493
    temperature_celsius: 26
    replicates: 2
  - step_description: Cells were cultured with pbs to facilitate down.
    conditions_or_variables:
    - 3 washes with lysis buffer
    data_collected: false
    temperature_celsius: 36
  - step_description: Cells were washed with sds-page loading buffer to facilitate
      say.
    conditions_or_variables:
    - 100V constant voltage
    - serum-free media
    data_collected: false
    duration_minutes: 634
    temperature_celsius: 18
control_groups:
- control_type: Isotype Control
  description: Their them explain who executive soon certain good or economic worker.
data_analysis_methods:
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
- One-way ANOVA with Tukey's post-hoc test
