How do you sterilize Arabidopsis seeds?

How do you sterilize Arabidopsis seeds?

Surface sterilization of Arabidopsis seeds

1. Put your seeds in a 1.7 ml tube or a falcon tube (depends on the amount of your seeds). Add enough 70% EtOH.
2. Pour out the EtOH.
3. Discard the bleach.
4. Wash seeds thoroughly with a large amount of sterile dH2O.
5. Discard the water.
6. Your seeds are ready for use.

Where can I buy Arabidopsis thaliana seeds?

You can get seeds from Arabidopsis Biological Resource Center (or ABRC; https://abrc.osu.edu/), at the Ohio State University, USA. If you just want some wild-type seeds, just ask some labs of regional research universities in India. The seeds of Arabidopsis according to your requirement can be procured from TAIR.

How do you sterilize seeds before planting?

Treating seeds with a solution of chlorine bleach can effectively remove bacterial pathogens and some viruses (such as tobacco mosaic virus) from the surfaces of seeds. 2. Add a drop or two of liquid dishwashing detergent or a commercial surfactant such as Activator 90 or Silwet to the disinfectant solution.

How do you sterilize seeds for tissue culture?

Swirl the flask containing the seed and bleach or repeatedly draw and aspirate the bleach solution in and out of the pipet. Sterilize the seed for 5-10 minutes. Remove the bleach solution and rinse the seed with sterile tissue culture grade water. Transfer the seed to sterile culture medium.

How does hydrogen peroxide disinfect seeds?

Use a 3% hydrogen peroxide solution and pour it into a small, watertight container. Leave the seeds for around four hours and this will kill off bacteria without affecting the seeds. You should then rinse the seeds thoroughly in clean, fresh water and then leave them to dry on a kitchen towel or a clean tea towel.

How do you sterilize seeds with hydrogen peroxide?

Soak your seeds in 3% hydrogen peroxide for 30 minutes. Rinse the seeds several times with water before planting and plant them as usual. Doing this breaks down the hard outer covering of the seeds and kills any pathogen present on them.

Which treatment is effective against surface sterilization of seed?

Seeds surface may sterilize with 70% ethanol for one minute followed by 6% sodium hypochlorite for 10 min. Then, the sterilized seeds have to wash by distilled water for 4–5 times.

How are Arabidopsis seeds sterilized?

Currently, Arabidopsis seeds are sterilized using two distinct sterilization techniques in conditions that differ slightly between labs and have not been standardized, often resulting in only partially effective sterilization or in excessive seed mortality.

How can I remove wax from Arabidopsis seeds?

I do a 1st wash with etanol 96%, for 1 min to remove wax and them the Mohammad Ali protocol. Simple surface sterilization of Arabidopsis seeds using 0.1% SDS for 5-6 minutes, 70% ethanol treatment for 2 minutes followed by five times wash with sterile distill H2O is effective in Arabidopsis and have worked all the times in our case.

How can I sterilize my seeds?

For A.th. seed sterilization, we use a 1-2 min wash with 70% ethanol, than the Mohammad Ali protocol, it works well! Seed aliquots are placed in clean 1.5 mL microcentrifuge tubes. Seeds are rinsed with 500 µl of 100% ethanol for 2 min. Mix well!

How do you germinate Arabidopsis seeds on MS plates?

Growth of Arabidopsis on MS plates Place the plates with the lid on top for three days at 4 ˚C and ambient humidity. Note: This process is called stratification and serves to synchronize the germination of individual seeds. Transfer the plates to the growth environment.