I want to use Alfalfa seeds for bio-remediation of oil contaminated soil and i want to sterilize them by using bleach. Any bleach that can i use it and for time. please i want references about that.
For doing surface sterilization I did usually using Chlorox or Bayclin that containing ca 5 % NaOCl; Generally I mixed Chlorox/Bayclin solutions with Ethanol 96 % 1: 5, v/v, incubation time 5-10 min, after incubation, than I washed the samples 3 times with sterilized water. All works were done in Laminar Air Flow cabinet. I think you can use any brand of whitening agents that containing NaOCl 5 %.
For surface sterilization, I also did at first by washing the explants with distilled water, ethanol 95 %, and surface sterilized using 1-2 % NaOCl (in water) for ca. 5 min, than wash three times with sterile water.
wash the seed with 70% ethanol 3 times and than wash with blech and after that wash with deionized water...
You can use any brand of bleach, but make sure it is not old and when you have to use it immediately after you make the solution because the bleach deactivated by light. the warm water mix with the bleach work very well. You may need to wash the seed with detergent first to dissolve the wax layer then wash them with the bleach mix .
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