Mimulus:Seed Sterilization: Difference between revisions
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#Immediately add 600 uL of Water. Do only 2 tubes at a time, because the alcohol treatment is harsh and might affect germination if is left on seeds too long | #Immediately add 600 uL of Water. Do only 2 tubes at a time, because the alcohol treatment is harsh and might affect germination if is left on seeds too long | ||
#Immediately suck off 600 uL of supernatant | #Immediately suck off 600 uL of supernatant | ||
#Repeat steps 3 and 4 four times | #Repeat steps 3 and 4 four times with water, not ethanol, to wash | ||
#Set aside tubes and repeat steps 1 to 5 for all the remaining tubes | #Set aside tubes and repeat steps 1 to 5 for all the remaining tubes | ||
#Add 200 uL of 20% bleach to each tube | #Add 200 uL of 20% bleach to each tube | ||
# | #Immediatly remove bleach solution | ||
#Add 600 uL of water | #Add 600 uL of water | ||
#Suck off 600 uL of the supernatant | #Suck off 600 uL of the supernatant | ||
#Repeat steps 9 and 10 four or five times | #Repeat steps 9 and 10 four or five times to wash | ||
Latest revision as of 13:14, 1 February 2007
Seed Sterilization
Supplies
- 70% Ethanol
- 20% Bleach
- Sterile Water
- Seeds
Methods
- Put seeds in an Eppendorf tube
- Add 200uL of 70% EtOH to tube
- Immediately add 600 uL of Water. Do only 2 tubes at a time, because the alcohol treatment is harsh and might affect germination if is left on seeds too long
- Immediately suck off 600 uL of supernatant
- Repeat steps 3 and 4 four times with water, not ethanol, to wash
- Set aside tubes and repeat steps 1 to 5 for all the remaining tubes
- Add 200 uL of 20% bleach to each tube
- Immediatly remove bleach solution
- Add 600 uL of water
- Suck off 600 uL of the supernatant
- Repeat steps 9 and 10 four or five times to wash
