Root Hair Cell Isolation


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).

  1. 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.
  2. 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.
  3. 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)
  4. 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.
  5. The LN2 containing root hair is then filtered through 90um sieve into 500ml glass beakers.
  6. After evaporation of LN2, the LN2 containing the root hairs is transferred into 50ml Falcon tube.
  7. When the LN2 is completely evaporated, the falcon tube with the root hair should store in -80°C for later usage.