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Seed Planting & Tools

 

 

 

 

              
  • 3 pieces of filter paper
  • Previously made agar plates with labels
  • Prepared seed stocks (RIL or NIL)
  • 100 µl pipette
  • Pipette tips
  • Seed sterilizing solution (70 % EtOH, 1% Triton X)
  • 70% EtOH
  • Planting template
  • Planting tool
  • RO water (Reverse Osmosis)
  • Black permanent marker
  • Microcentrifuge tubes
  • Rubberbands

Get a complete listing of tools

5) Planting Seeds

5.1)  Place 3 pieces of filter paper on workspace, check plate label for appropriate seed numbers, and mark each filter paper with the appropriate seed number.
5.2) Find those seed line numbers on a printed list of seed numbers, and determine how many of each seed are needed. Note: For this experiment, three seeds are used for each designated seed line. e.g. if there are three number 34's on the printed list, 9 seeds from tube 34 are needed.
5.3) Pour the correct amount of seed on the coinciding filter paper
5.4) Using a 100 microliter pipette, saturate the seeds with seed sterilizer (70% EtOH, 1% Triton X), then, using a different tip, cover them with 70% ethanol. 
5.5) Line up the labeled agar plate on designated template.
5.6) Dip planting tool in microcentrifuge tube filled with water, gently pick up specified seed, and place on the surface of the agar. Note: be very careful with this step as we don't want to damage the agar in any way. 
5.7) Make sure to line the seeds up exactly with the pen dots on the template. It's also very important to pay attention to the seed numbers on the label and the order you're placing them on the surface of the agar. The seed lines should match up with those on the label. 
5.8) After all 9 seeds are planted, make a vertical black line on the outside of the agar plate (use the template as a guide) using permanent marker. 
5.9) Place the lid on the agar plate and secure the bottom of the petri dish by wrapping parafilm around the edge of the plate. 
5.10) If any extra seeds were sterilized for future plates, place them in separate designated microcentrifuge tubes. e.g. If the filter paper labeled 34 has 6 extra seeds, place them in a separate tube for sterilized seeds labeled 34 small or large. If there is no tube with that number, make a new one. Pay attention to whether you're working with small seeds or large seeds. 
5.11) Repeat steps 5.1-5.10 with each new agar plate and label. 
5.12) When all plates for that day have been planted, stack them numerically in groups of 6.
5.13) Place the plates in the refrigerator on the day designated by the label, and let stratify for 5 days.