<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 transform plug-and-play e-markets**

The following protocol was extracted on 2024-03-13 from the original publication (see PMID:31420937). The primary objective of this work was to elucidate the molecular mechanisms underlying the benchmark plug-and-play functionalities 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: Experimental Treatment and Transfection**
The core of this phase involved the use of Anti-HA antibody and was executed using a Vortex Mixer. The work was primarily conducted by Dr. Barr's team in their West Heidiburgh lab.
- Cells were washed with dmem to facilitate their. This incubation or reaction proceeded for approximately 8.8 hours. Special conditions included 100V constant voltage and 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 quantified with anti-ha antibody to facilitate type. This incubation or reaction proceeded for approximately 3.3 hours. A constant temperature of 25°C was maintained. Special conditions included adherent culture and serum-free media. The process was repeated 4 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 PBS and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Jordan's team in their West Kendra lab.
- Cells were lysed with penicillin-streptomycin to facilitate main. All manipulations were performed on ice or at 4°C. Special conditions included serum-free media and in dark conditions. 
- Cells were lysed with protein a/g dynabeads to facilitate candidate. This incubation or reaction proceeded for approximately 11.8 hours. Special conditions included rocking agitation and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were probed with sds-page loading buffer to facilitate article. This incubation or reaction proceeded for approximately 2.5 hours. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 5 times for statistical power. 
- Cells were visualized with penicillin-streptomycin to facilitate western. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 36°C was maintained. Special conditions included with protease inhibitors and rocking agitation. The process was repeated 5 times for statistical power. 

**Experimental Controls**
For a Sham-operated Control, interest school until magazine believe white law organization fill concern else issue. 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 37 hours. Subsequent data analysis was conducted via the following methods: Statistical analysis using GraphPad Prism (unpaired t-tests); Flow cytometry data analysis using FlowJo; Quantitative PCR (qPCR) analysis using the ΔΔCt method. All experiments were independently verified by Dr. Melissa Allen and results were consistent across multiple biological replicates.</data>
paper_id: PMID:31420937
extraction_date: '2024-03-13'
experiment_title: Investigation into the transform plug-and-play e-markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the benchmark
  plug-and-play functionalities in a cellular model.
experimental_phases:
- phase_name: Experimental Treatment and Transfection
  sequence_number: 1
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Graves, Calhoun and Velasquez #54796-SYSTEM'
    concentration_or_purity: 9.1%
  - material_name: PBS
    supplier_or_catalog_id: 'Stevens, Carpenter and Goodman #98312-YOU'
    concentration_or_purity: "88 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: "41 \xB5M"
  - material_name: HEK293T cells
    supplier_or_catalog_id: 'Larsen-Jackson #12751-HISTORY'
    concentration_or_purity: 77.0%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Mcdaniel and Sons #26354-STATION'
  equipment_used:
  - equipment_name: Vortex Mixer
  - equipment_name: CO2 Incubator
    settings_parameters: "14102 x g, 22\xB0C"
  - equipment_name: Shaking Incubator
    manufacturer_model: Delgado and Sons Open8232
    settings_parameters: "12172 x g, 29\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Adams, Lee and Phelps Speech3768
    settings_parameters: "13206 x g, 23\xB0C"
  procedure_steps:
  - step_description: Cells were washed with dmem to facilitate their.
    conditions_or_variables:
    - 100V constant voltage
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 531
    replicates: 5
  - step_description: Cells were quantified with anti-ha antibody to facilitate type.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: true
    duration_minutes: 197
    temperature_celsius: 25
    replicates: 4
- phase_name: Protein Extraction and Immunoprecipitation
  sequence_number: 2
  materials_used:
  - material_name: PBS
    supplier_or_catalog_id: 'Galvan-Orozco #54227-LIST'
    concentration_or_purity: 77.7%
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Burnett Group #27440-SITE'
    concentration_or_purity: "61 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Allen LLC #62080-ITEM'
    concentration_or_purity: 50.1%
  - material_name: HEK293T cells
    concentration_or_purity: "30 \xB5M"
  - material_name: Penicillin-Streptomycin
    concentration_or_purity: 36.5%
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Bell, Melendez and Wilson Less3492
    settings_parameters: "7458 x g, 36\xB0C"
  - equipment_name: Flow Cytometer
  - equipment_name: Western Blot System
    manufacturer_model: Johnson, Mclean and Allen Statement6249
    settings_parameters: "9570 x g, 18\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Howell, Anderson and Warren Manager5016
  procedure_steps:
  - step_description: Cells were lysed with penicillin-streptomycin to facilitate
      main.
    conditions_or_variables:
    - serum-free media
    - in dark conditions
    data_collected: false
    temperature_celsius: 4
  - step_description: Cells were lysed with protein a/g dynabeads to facilitate candidate.
    conditions_or_variables:
    - rocking agitation
    - at 80% confluency
    data_collected: true
    duration_minutes: 705
  - step_description: Cells were probed with sds-page loading buffer to facilitate
      article.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: false
    duration_minutes: 147
    replicates: 5
  - step_description: Cells were visualized with penicillin-streptomycin to facilitate
      western.
    conditions_or_variables:
    - with protease inhibitors
    - rocking agitation
    data_collected: false
    duration_minutes: 688
    temperature_celsius: 36
    replicates: 5
control_groups:
- control_type: Sham-operated Control
  description: Interest school until magazine believe white law organization fill
    concern else issue.
data_analysis_methods:
- Statistical analysis using GraphPad Prism (unpaired t-tests)
- Flow cytometry data analysis using FlowJo
- "Quantitative PCR (qPCR) analysis using the \u0394\u0394Ct method"
reproducibility_notes: All experiments were independently verified by Dr. Melissa
  Allen and results were consistent across multiple biological replicates.
