Root Hair Isolation Steps:
The following steps are performed in liquid nitrogen (LN2). The average liquid nitrogen consumption is 5L liquid nitrogen/ sample (30-60 plants).
- The root systems of soybean/sorghum plants are collected, rapidly wiped off to remove extra moisture then cut the root off and immediately immersed into liquid nitrogen.
- Frozen roots are gently stirred into liquid nitrogen by a glass rod for 20 minutes. The flow of liquid nitrogen is sufficient to break root and root hairs.
- The liquid nitrogen containing the root hairs is filtered through 500μm sieve into a 2000 ml glass beaker. (Can use funnel to connect the sieve and beaker)
- The stripped roots are put back into the LN2 tank and rinsed 5 to 7 times with LN2 to collect the remaining root hair cells and increase the yield.
- The LN2 containing root hair is then filtered through 90um sieve into 500ml glass beakers.
- After evaporation of LN2, the LN2 containing the root hairs is transferred into 50ml Falcon tube.
- When the LN2 is completely evaporated, the falcon tube with the root hair should store in -80°C for later usage.