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Chemical SOPs

University of Washington Lab Safety Manual

Chemical Material Safety Data Sheets (MSDS) on MyChem website.

UW Hazardous Waste Online Collection Request

RNAzol RT

Purpose: RNAzol RT is used for separating RNA from cells.

Specific Hazards: RNAzol RT is corrosive and harmful if inhaled.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves

    • Chemical fume hood

  • WHEN SPLASH POTENTIAL EXISTS

    • Safety goggles
    • Lab Coat

Waste Disposal:

  • Liquid Waste:
    • Transfer liquid waste to labeled container designated for RNAzol waste.
  • Solid Waste:
    • Tubes/tip/gloves with residual RNAzol should be stored in chemical fume hood for no less than 24hrs and then disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Isopropanol (2-propanol)

Purpose: Used in the precipitation of nucleic acids.

Specific Hazards: Highly flammable - keep away from ignition sources.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves

  • WHEN SPLASH POTENTIAL EXISTS

    • Safety goggles

    • Lab Coat

Waste Disposal:

  • Liquid Waste:

    • Transfer liquid waste to labeled container designated for isopropanol waste.

  • Solid Waste:

    • Tubes/tip/gloves with residual isopropanol should be stored in chemical fume hood for no less than 24hrs and then disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Diethylpyrocarbonate (DEPC)

Purpose: Used as an RNase inhibitor during handling and storage of RNA.

Specific Hazards:

  • Skin irritant and harmful if inhaled.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves
    • Chemical fume hood

  • WHEN SPLASH POTENTIAL EXISTS

    • Safety goggles
    • Lab Coat

Waste Disposal:

  • Liquid Waste:
    • Transfer liquid waste to labeled container designated for DEPC waste.
  • Solid Waste:
    • Tubes/tip/gloves with residual DEPC should be stored in chemical fume hood for no less than 24hrs and then disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

DNAzol

Purpose: DNAzol is used for separating DNA from cells.

Specific Hazards:

  • DNAzol is corrosive.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves

  • WHEN SPLASH POTENTIAL EXISTS

    • Safety goggles
    • Lab Coat

Waste Disposal:

  • Liquid Waste:
    • Transfer liquid waste to labeled container designated for DNAzol waste.
  • Solid Waste:
    • Tubes/tip/gloves with residual DNAzol should be stored in chemical fume hood for no less than 24hrs and then disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Ethanol

Purpose: Used in the precipitation of nucleic acids.

Specific Hazards:

  • Highly flammable - keep away from ignition sources.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves

  • WHEN SPLASH POTENTIAL EXISTS

    • Safety goggles
    • Lab Coat

Waste Disposal:

  • Liquid Waste:

    • Transfer liquid waste to labeled container designated for ethanol waste.

  • Solid Waste:

    • Tubes/tip/gloves with residual ethanol should be stored in chemical fume hood for no less than 24hrs and then disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Phenol:chloroform:IAA (25:24:1)

Purpose: Used for purification of nucleic acids.

Specific Hazards:

  • Phenol:chloroform:IAA is corrosive and harmful if inhaled.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves
    • Chemical fume hood

  • WHEN SPLASH POTENTIAL EXISTS

    • Safety goggles
    • Lab Coat

Waste Disposal:

  • Liquid Waste:

    • Transfer liquid waste to labeled container designated for phenol:chloroform:IAA waste.

  • Solid Waste:

    • Tubes/tip/gloves with residual phenol:chloroform:IAA should be stored in chemical fume hood for no less than 24hrs and then disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Proteinase K

Purpose: Enzyme used to degrade proteins during DNA isolation.

Specific Hazards:

  • None.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves

Waste Disposal:

  • No requirements.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Resazurin assay

Personal Protective Equipment:

  • REQUIRED:

    • Gloves

Chemical exposures:

  • dimethyl sulfoxide (DMSO)

  • flammable

  • skin irritant

  • HyClone antibiotic antimycotic (PEN/STREP/FUNGIZONE) solution

  • skin irritant

  • resazurin

Waste Disposal:

  • Dispose of all waste in labelled resazurin waste container.

  • Plate oysters (1/well; if testing in 2-3 mm spat one can use a 96 well plate and 300 ul of assay media. If using larger oysters you can use a 48 well plate and 1 mL of assay media.)
    When you plate them you can do so in seawater with antibiotics and fungizone. Weigh oysters as you plate them so you can correct for weight. Alternatively, plate them, take a picture and measure area using image J.

  • Seawater: (Volume/100 mL)

    1. Sterile seawater with appropriate Salinity – 99 mL (Sterile Filter or Autoclave)
    2. 100X Penn/Strep/Fungizone – 1 mL

  • Allow oysters to be in Antibiotic/Fungizone seawater for at least 1 h

  • Make Resazurin Stock Solution (This can be frozen; 50 mg/ml)

  • 0.5 g of resazurin sodium salt (Sigma-Aldrich Corp., St. Louis, MO)
  • 10 mL of distilled water

  • 10 μL of dimethyl sulfoxide (0.1% DMSO, Sigma-Aldrich Corp., St. Louis, MO)

  • Make Assay Media

  • Sterile seawater with appropriate Salinity – 97.9 mL (Sterile Filter or Autoclave)
  • Resazurin Stock Solution – 1 mL (This can be replaced with AlamarBlue, which is used at 1.6%)
  • 100X Penn/Strep/Fungizone – 1 mL

  • DMSO – 100 uL

  • Set Gain on Plate Reader

  • Plate 300 ul into blank well of 96 well plate and read (Excitation 530; Emission 590)

  • Adjust gain manually until signal is around 500. This is essential to ensure sensitivity.

  • Add Assay Media to each well (300 ul in 96-well plate or 1 mL in 48 well plate).

  • Read plate immediately upon adding Assay media

  • Read plate at intervals thereafter (if you are at 30 ppt this should occur at 1, 3, and 6 h; if you are at 4 ppt you may want 6, 12, and 24h).
    This allows you to show that signal increased linearly with time. If you find that you have a specific timepoint that you like for future studies you can just use that timepoint.

  • Calculate the change in fluorescence.

  • Calculate the change in fluorescence/area or mass.

  • Sort oysters according to a high or low change in fluorescence/area or mass (low change will grow more quickly).

Prepared by:

  • Name: Sam White
  • Date: 2024-07-19

Sodium acetate

Purpose: Used to precipitate nucleic acids.

Specific Hazards:

  • Skin irritant.

Personal Protective Equipment:

  • REQUIRED:
    • Gloves

Waste Disposal:

  • Liquid Waste:

    • Dispose in sink.

  • Solid Waste:

    • Tubes/tip/gloves with residual sodium acetate can be disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Ethidium bromide (EtBr)

Purpose: Use for staining nucleic acids in agarose gels.

Specific Hazards:

  • Mutagen and potential carcinogen.

Personal Protective Equipment:

  • REQUIRED:

    • Gloves

  • WHEN SPLASH POTENTIAL EXISTS

    • Safety goggles
    • Lab Coat

Waste Disposal:

NOTE: Disposal instructions are only for dilute EtBr concentrations (< 10ug/mL) that are used in the lab. Concentrations higher than this are hazardous waste and should be placed in a labeled, dedicated container.

  • Liquid Waste:

    • Dispose in sink.

  • Solid Waste:

    • Tubes/tip/gloves/gels should be double bagged, labelled "non-hazardous", and then disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24

Qiagen Kits

Purpose: Use for isolation of nucleic acids.

Specific Hazards:

  • Harmful, irritant.

NOTE: Do not combine kit reagents with bleach (sodium hypchlorite)!

Personal Protective Equipment:

  • REQUIRED:
    • Gloves
  • WHEN SPLASH POTENTIAL EXISTS
    • Safety goggles
    • Lab Coat

Waste Disposal:

  • Liquid Waste:

    • Transfer liquid waste to labeled container designated for Qiagen Kit waste.

  • Solid Waste:

    • Tubes/tip/gloves with residual sodium acetate can be disposed in regular trash.

Prepared by:

  • Name: Sam White
  • Date: 2016-10-24