Preparing spore samples
Obvious Reminder: DO NOT contaminate or mix samples at any step
1. Wipe down a hood with 70% ethanol – do not turn on flow vents
2. Get together some forceps, a glass rod, a Bunsen burner, a beaker with 95% ethanol,
KimWipes, eppendorf tubes, and Triton water (1 drop Triton in 100 mL ddH2O)
a. Make sure the Bunsen burner and beaker are AT LEAST a foot apart to avoid
igniting the fumes from the ethanol
3. Number and fill an eppendorf tube for each sample with 200 µL of Triton water
a. You can also use an 8-tube strip if you are working with many samples. Tape up
wells that you are not using – samples will be dropped and it is better to drop
them on tape rather than in another well.
4. Take your sample (flower) out of the coin envelope and place it on the KimWipe
5. Use the forceps to carefully remove a single smutted anther and place it into a tube
a. While anthers are the best, you can also use any smutted floral tissues
6. Record the tube number and the sample that you just used on a datasheet
7. Repeat steps 4–6, making sure to use new KimWipes and sterilize with ethanol and fire
between samples
8. It is possible for spores to germinate while suspended in solution, so do not leave them in
eppendorf tubes for more than 24 hours before using them.