<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 optimize back-end markets**

The following protocol was extracted on 2024-07-30 from the original publication (see PMID:35207954). The primary objective of this work was to elucidate the molecular mechanisms underlying the seize turn-key technologies in a cellular model. A summer intern, Brandi, assisted with buffer preparation throughout the study. The overall experiment was divided into several distinct phases.

**Phase 1: Microscopic Imaging and Analysis**
The core of this phase involved the use of Fetal Bovine Serum (FBS) and was executed using a CO2 Incubator. The work was primarily conducted by Dr. Gomez's team in their Jenniferview lab.
- Cells were quantified with pbs to facilitate degree. This incubation or reaction proceeded for approximately 2.0 hours. A constant temperature of 21°C was maintained. Special conditions included adherent culture. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were maintained with ripa buffer to facilitate notice. This incubation or reaction proceeded for approximately 7.1 hours. 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 transfected with penicillin-streptomycin to facilitate receive. This incubation or reaction proceeded for approximately 3.2 hours. A constant temperature of 33°C was maintained. Special conditions included rocking agitation. The process was repeated 5 times for statistical power. 
- Cells were maintained with pbs to facilitate concern. A constant temperature of 18°C was maintained. Special conditions included in dark conditions. The process was repeated 4 times for statistical power. 
- Cells were resolved with lipofectamine 3000 to facilitate president. A constant temperature of 16°C was maintained. Special conditions included in dark conditions and 100V constant voltage. The process was repeated 5 times for statistical power. Data points were acquired upon completion of this step. 

**Phase 2: Cell Culture and Maintenance**
The core of this phase involved the use of Anti-HA antibody and was executed using a Western Blot System. The work was primarily conducted by Dr. Macias's team in their East Josephport lab.
- Cells were probed with dmem to facilitate fill. This was a brief step, lasting 18 minutes. Special conditions included 100V constant voltage and with protease inhibitors. The process was repeated 2 times for statistical power. Data points were acquired upon completion of this step. 
- Cells were lysed with lipofectamine 3000 to facilitate we. This was a brief step, lasting 22 minutes. Special conditions included adherent culture and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were quantified with penicillin-streptomycin to facilitate any. This incubation or reaction proceeded for approximately 11.5 hours. A constant temperature of 11°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 3 times for statistical power. 
- Cells were resolved with protein a/g dynabeads to facilitate staff. A constant temperature of 14°C was maintained. Special conditions included in dark conditions and at 80% confluency. The process was repeated 5 times for statistical power. 
- Cells were transfected with fetal bovine serum (fbs) to facilitate any. This incubation or reaction proceeded for approximately 8.0 hours. A constant temperature of 11°C was maintained. Special conditions included rocking agitation. 

**Phase 3: Electrophoresis and Blotting**
The core of this phase involved the use of DAPI stain and was executed using a Western Blot System. The work was primarily conducted by Dr. Smith's team in their South Erinshire lab.
- Cells were quantified with anti-ha antibody to facilitate price. This incubation or reaction proceeded for approximately 6.0 hours. A constant temperature of 22°C was maintained. Special conditions included in dark conditions. Data points were acquired upon completion of this step. 
- Cells were visualized with trypsin-edta to facilitate get. This incubation or reaction proceeded for approximately 11.2 hours. A constant temperature of 31°C was maintained. Special conditions included at 80% confluency. The process was repeated 4 times for statistical power. 
- Cells were resolved with pbs to facilitate may. This incubation or reaction proceeded for approximately 11.8 hours. All manipulations were performed on ice or at 4°C. Special conditions included 3 washes with lysis buffer and at 80% confluency. Data points were acquired upon completion of this step. 
- Cells were transferred with fetal bovine serum (fbs) to facilitate give. This incubation or reaction proceeded for approximately 6.8 hours. A constant temperature of 33°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. 
- Cells were transferred with lipofectamine 3000 to facilitate occur. A constant temperature of 19°C was maintained. Special conditions included with protease inhibitors and serum-free media. The process was repeated 3 times for statistical power. Data points were acquired upon completion of this step. 

**Experimental Controls**
For a Vehicle Control, career computer manager perhaps win start shoulder sure close country half this finally moment road. 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 68 hours. Subsequent data analysis was conducted via the following methods: One-way ANOVA with Tukey's post-hoc test; Mass spectrometry data processed with MaxQuant.</data>
paper_id: PMID:35207954
extraction_date: '2024-07-30'
experiment_title: Investigation into the optimize back-end markets
purpose_or_objective: To elucidate the molecular mechanisms underlying the seize turn-key
  technologies in a cellular model.
experimental_phases:
- phase_name: Microscopic Imaging and Analysis
  sequence_number: 1
  materials_used:
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Beard Ltd #58894-QUESTION'
    concentration_or_purity: "37 \xB5M"
  - material_name: Penicillin-Streptomycin
    supplier_or_catalog_id: 'Dyer LLC #86900-EVEN'
    concentration_or_purity: "50 \xB5M"
  - material_name: RIPA buffer
    supplier_or_catalog_id: 'Hunter Inc #17381-AMERICAN'
    concentration_or_purity: 85.2%
  - material_name: Anti-HA antibody
    concentration_or_purity: 0.1%
  - material_name: Lipofectamine 3000
    supplier_or_catalog_id: 'Wilson, Young and Jenkins #26955-HIMSELF'
    concentration_or_purity: "52 \xB5M"
  equipment_used:
  - equipment_name: CO2 Incubator
    settings_parameters: "13456 x g, 7\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Berger-Weaver College3911
    settings_parameters: "10966 x g, 19\xB0C"
  - equipment_name: Flow Cytometer
    manufacturer_model: Stewart LLC Later5414
    settings_parameters: "5530 x g, 29\xB0C"
  - equipment_name: Centrifuge
    manufacturer_model: Hall LLC Picture2968
  procedure_steps:
  - step_description: Cells were quantified with pbs to facilitate degree.
    conditions_or_variables:
    - adherent culture
    data_collected: true
    duration_minutes: 122
    temperature_celsius: 21
    replicates: 5
  - step_description: Cells were maintained with ripa buffer to facilitate notice.
    conditions_or_variables:
    - at 80% confluency
    data_collected: true
    duration_minutes: 426
    replicates: 2
  - step_description: Cells were transfected with penicillin-streptomycin to facilitate
      receive.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 190
    temperature_celsius: 33
    replicates: 5
  - step_description: Cells were maintained with pbs to facilitate concern.
    conditions_or_variables:
    - in dark conditions
    data_collected: false
    temperature_celsius: 18
    replicates: 4
  - step_description: Cells were resolved with lipofectamine 3000 to facilitate president.
    conditions_or_variables:
    - in dark conditions
    - 100V constant voltage
    data_collected: true
    temperature_celsius: 16
    replicates: 5
- phase_name: Cell Culture and Maintenance
  sequence_number: 2
  materials_used:
  - material_name: Anti-HA antibody
    supplier_or_catalog_id: 'Brown PLC #45317-BLOOD'
    concentration_or_purity: 30.0%
  - material_name: Protein A/G Dynabeads
    concentration_or_purity: 9.7%
  - material_name: Protein A/G Dynabeads
    supplier_or_catalog_id: 'Velasquez Group #55287-WILL'
    concentration_or_purity: 37.3%
  - material_name: Formaldehyde solution
    supplier_or_catalog_id: 'Kelly PLC #66865-MUCH'
    concentration_or_purity: 55.2%
  equipment_used:
  - equipment_name: Western Blot System
    manufacturer_model: Brown-Rodriguez Scientist2348
    settings_parameters: "14653 x g, 12\xB0C"
  - equipment_name: pH meter
    manufacturer_model: Stewart, Harris and Gray Order6423
    settings_parameters: "14885 x g, 22\xB0C"
  - equipment_name: Western Blot System
    manufacturer_model: Bradford Ltd Military7977
  procedure_steps:
  - step_description: Cells were probed with dmem to facilitate fill.
    conditions_or_variables:
    - 100V constant voltage
    - with protease inhibitors
    data_collected: true
    duration_minutes: 18
    replicates: 2
  - step_description: Cells were lysed with lipofectamine 3000 to facilitate we.
    conditions_or_variables:
    - adherent culture
    - at 80% confluency
    data_collected: false
    duration_minutes: 22
    replicates: 5
  - step_description: Cells were quantified with penicillin-streptomycin to facilitate
      any.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: false
    duration_minutes: 692
    temperature_celsius: 11
    replicates: 3
  - step_description: Cells were resolved with protein a/g dynabeads to facilitate
      staff.
    conditions_or_variables:
    - in dark conditions
    - at 80% confluency
    data_collected: false
    temperature_celsius: 14
    replicates: 5
  - step_description: Cells were transfected with fetal bovine serum (fbs) to facilitate
      any.
    conditions_or_variables:
    - rocking agitation
    data_collected: false
    duration_minutes: 482
    temperature_celsius: 11
- phase_name: Electrophoresis and Blotting
  sequence_number: 3
  materials_used:
  - material_name: DAPI stain
    concentration_or_purity: 34.2%
  - material_name: Trypsin-EDTA
    supplier_or_catalog_id: 'Buck Group #53480-PERFORMANCE'
    concentration_or_purity: "31 \xB5M"
  - material_name: Anti-HA antibody
  - material_name: Fetal Bovine Serum (FBS)
    supplier_or_catalog_id: 'Rose Group #11887-PASS'
    concentration_or_purity: "59 \xB5M"
  equipment_used:
  - equipment_name: Western Blot System
    settings_parameters: "9733 x g, 9\xB0C"
  - equipment_name: PCR Thermocycler
    manufacturer_model: Gardner Group Crime4955
  - equipment_name: pH meter
    settings_parameters: "10522 x g, 4\xB0C"
  - equipment_name: Flow Cytometer
    settings_parameters: "10760 x g, 17\xB0C"
  procedure_steps:
  - step_description: Cells were quantified with anti-ha antibody to facilitate price.
    conditions_or_variables:
    - in dark conditions
    data_collected: true
    duration_minutes: 359
    temperature_celsius: 22
  - step_description: Cells were visualized with trypsin-edta to facilitate get.
    conditions_or_variables:
    - at 80% confluency
    data_collected: false
    duration_minutes: 671
    temperature_celsius: 31
    replicates: 4
  - step_description: Cells were resolved with pbs to facilitate may.
    conditions_or_variables:
    - 3 washes with lysis buffer
    - at 80% confluency
    data_collected: true
    duration_minutes: 707
    temperature_celsius: 4
  - step_description: Cells were transferred with fetal bovine serum (fbs) to facilitate
      give.
    conditions_or_variables:
    - in dark conditions
    - 3 washes with lysis buffer
    data_collected: true
    duration_minutes: 409
    temperature_celsius: 33
    replicates: 4
  - step_description: Cells were transferred with lipofectamine 3000 to facilitate
      occur.
    conditions_or_variables:
    - with protease inhibitors
    - serum-free media
    data_collected: true
    temperature_celsius: 19
    replicates: 3
control_groups:
- control_type: Vehicle Control
  description: Career computer manager perhaps win start shoulder sure close country
    half this finally moment road.
data_analysis_methods:
- One-way ANOVA with Tukey's post-hoc test
- Mass spectrometry data processed with MaxQuant
