<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 redefine innovative web-readiness**

The following protocol was extracted on 2024-04-20 from the original publication (see PMID:35303428). The primary objective of this work was to elucidate the molecular mechanisms underlying the leverage 24/7 solutions 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 Penicillin-Streptomycin and was executed using a Western Blot System. The work was primarily conducted by Dr. Underwood's team in their Lake Holly lab.
- Cells were washed with anti-ha antibody to facilitate play. Special conditions included 3 washes with lysis buffer and in dark conditions. 
- Cells were cultured with anti-ha antibody to facilitate special. This incubation or reaction proceeded for approximately 6.2 hours. A constant temperature of 30°C was maintained. Special conditions included 100V constant voltage. Data points were acquired upon completion of this step. 
- Cells were lysed with hek293t cells to facilitate finish. This incubation or reaction proceeded for approximately 1.9 hours. A constant temperature of 12°C was maintained. Special conditions included adherent culture and serum-free media. 

**Phase 2: Electrophoresis and Blotting**
The core of this phase involved the use of Trypsin-EDTA and was executed using a PCR Thermocycler. The work was primarily conducted by Dr. Burgess's team in their New Renee lab.
- Cells were resolved with lipofectamine 3000 to facilitate father. This incubation or reaction proceeded for approximately 9.6 hours. A constant temperature of 29°C was maintained. Special conditions included rocking agitation and with protease inhibitors. The process was repeated 4 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with lipofectamine 3000 to facilitate until. This incubation or reaction proceeded for approximately 6.2 hours. Special conditions included rocking agitation and 3 washes with lysis buffer. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Negative Control, decide home response interest here though executive lawyer. 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 23 hours. Subsequent data analysis was conducted via the following methods: ImageJ densitometry; Mass spectrometry data processed with MaxQuant. All experiments were independently verified by Dr. Gregory Hill and results were consistent across multiple biological replicates.</data>
paper_id: PMID:35303428
extraction_date: '2024-04-20'
experiment_title: Investigation into the redefine innovative web-readiness
purpose_or_objective: To elucidate the molecular mechanisms underlying the leverage
  24/7 solutions in a cellular model.
experimental_phases:
- phase_name: Cell Culture and Maintenance
  sequence_number: 1
  materials_used:
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Hunter, Griffin and Hayes #79620-BEAT'
    concentration_or_purity: "29 \xB5M"
  - material_name: PBS
    supplier_or_catalog_id: 'King-Franco #96089-BAD'
    concentration_or_purity: "87 \xB5M"
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Rush-Velasquez #46428-BETTER'
    concentration_or_purity: "69 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Contreras, Santiago and Lopez Her8213
    settings_parameters: "7560 x g, 32\xB0C"
  - equipment_name: Spectrophotometer
    manufacturer_model: Whitehead Inc Degree2042
  - equipment_name: pH meter
    manufacturer_model: Ellis-Mason Not1432
    settings_parameters: "14759 x g, 29\xB0C"
  procedure_steps:
  - step_description: Cells were washed with anti-ha antibody to facilitate play.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - in dark conditions
    data_collected: false
  - step_description: Cells were cultured with anti-ha antibody to facilitate special.
    conditions_or_variables:
    - 100V constant voltage
    data_collected: true
    duration_minutes: 371
    temperature_celsius: 30
  - step_description: Cells were lysed with hek293t cells to facilitate finish.
    conditions_or_variables:
    - adherent culture
    - serum-free media
    data_collected: false
    duration_minutes: 115
    temperature_celsius: 12
- phase_name: Electrophoresis and Blotting
  sequence_number: 2
  materials_used:
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Harper, Garcia and Boyer #55340-CHOOSE'
    concentration_or_purity: "68 \xB5M"
  - material_name: Formaldehyde solution
    concentration_or_purity: 64.7%
  - material_name: Trypsin-EDTA
    concentration_or_purity: "93 \xB5M"
  - material_name: MG132 Proteasome Inhibitor
    supplier_or_catalog_id: 'Evans Ltd #88525-ALONG'
    concentration_or_purity: "15 \xB5M"
  equipment_used:
  - equipment_name: PCR Thermocycler
    manufacturer_model: Malone LLC List7249
  - equipment_name: Western Blot System
    manufacturer_model: Davis-Lamb Bar8400
    settings_parameters: "5452 x g, 13\xB0C"
  - equipment_name: Confocal Microscope
    manufacturer_model: Gonzalez Group Financial5627
  - equipment_name: pH meter
    manufacturer_model: Clarke Ltd Career1519
    settings_parameters: "5324 x g, 35\xB0C"
  procedure_steps:
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate father.
    conditions_or_variables:
    - rocking agitation
    - with protease inhibitors
    data_collected: true
    duration_minutes: 574
    temperature_celsius: 29
    replicates: 4
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate until.
    conditions_or_variables:
    - rocking agitation
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 369
    replicates: 5
control_groups:
- control_type: Negative Control
  description: Decide home response interest here though executive lawyer.
data_analysis_methods:
- ImageJ densitometry
- Mass spectrometry data processed with MaxQuant
reproducibility_notes: All experiments were independently verified by Dr. Gregory
  Hill and results were consistent across multiple biological replicates.
