Standard Operating Procedure: Arabidopsis Tissue Sampling
Title:
Department: Agronomy
Created by: Danielle Dykema, Kendra Meade
Laboratory: Crop Production & Physiology Lab Suite
Supervisor: Bill Beavis
Lab Supervisor: Whitney Bouma
Date approved:
Procedure Overview:
This procedure describes the sampling of Arabidopsis thaliana tissue and cryopreservation. A
leaf cutting is taken from each plant and is then frozen in microcentrifuge tubes using liquid
nitrogen. The samples are then stored at -80 degrees Celsius for future use.
Equipment and reagents necessary:
Reagents:
Liquid Nitrogen
50mL 75% ethanol
Equipment:
Lab coat
Safety glasses
Sterile 1.5 mL microcentrifuge tubes
Microcentrifuge tube rack
-80ºC freezer
Permanent marker
Foreceps
Styrofoam container or ice bucket – minimum two quart size
Disposables:
50mL Falcon tube
Kemwipes
Procedure:
1. Place sterile tubes (closed) in microcentrifuge tube rack, label each with an individual plant
number
2. Pour liquid nitrogen into Styrofoam container or ice bucket until ½ full
3. Clean forceps with 75% ethanol solution, remove 1-2 leaves from a single plant and place in
the appropriate microcentrifuge tube
4. Close cap on microcentrifuge tube and place immediately into the liquid nitrogen
5. Sterilize forceps with 75% ethanol solution and repeat for all samples
6. Remove samples from the liquid nitrogen and place into microcentrifuge tube box
7. Store in -80°C freezer
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8. Allow liquid nitrogen to evaporate under fume hood while person running procedure is in
the lab
Personal Protective Equipment / Engineering Controls:
Lab coat, eye protection and hand protection must be selected as required by Section D of the EH&S
Laboratory Safety Manual. Check the box(es) next to the item(s) which are appropriate for your
procedure.
Nitrile gloves
Safety glasses
Face shield
Dust mask
Latex gloves
Splash goggles
Lab coat
Fume hood
Neoprene gloves
Vented goggles
Apron
Biosafety cabinet
Insulated gloves
Eye wash station
Safety shower
Respirator
Note: Open-toed and heeled shoes are NOT allowed.
Other Control Measures:
Handling & Storage Precautions:
High pressure gas. Do not puncture or incinerate container. Use equipment rated for cylinder pressure.
Close valve after each use and when empty. Protect cylinders from physical damage; do not drag, roll,
slide, or drop. Use a suitable hand truck for cylinder movement.
Never allow any unprotected part of the body to touch uninsulated pipes or vessels that contain cryogenic
liquids. Prevent entrapment of liquid in closed systems or piping without pressure relief devices. Some
materials may become brittle at low temperatures and will easily fracture.
Waste Disposal Procedures:
Dispose of in a manner consistent with federal, state, and local regulations.
Spill/Release Containment and Clean Up/Decontamination Procedures:
Immediately contact emergency personnel. Stop leak if without risk
Health & Safety Summary for Required Reagents:
Brief description of associated hazards
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 The above summary consists of guidelines for proper handling & disposal of
chemicals used in this procedure. You must read and understand the contents
of the entire MSDS(s) before starting this procedure.
References:
List references used in preparing this procedure.
revised 18 June 2008 me3