Description
Seed Sterilization and Germination Buffer B is corrosive. Please take effective measures to avoid inhalation or direct contact with skin. This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Instructions for Use：1.Seed preparation: Place 50-100 Arabidopsis or tobacco seeds in a 1.5ml centrifuge tube and proceed with subsequent steps in a Laminar Flow Cabinet. 2.Seed treatment with ethanol: Add 1ml of 70% ethanol to the centrifuge tube, invert the tube for 2 min and then place the tube on a plastic rack to allow seeds to settle at the bottom of the tube. Carefully remove the ethanol with a 1ml pipette. 3.Sterilization and germination: Add 250µl of Seed Sterilization and Germination Buffer A and 250µl of Seed Sterilization and Germination Buffer B to the seeds in sequence, mix well and soak the seeds for 1min (during which the seeds should be fully suspended to ensure sufficient sterilization). Allow seeds to settle at the bottom of the tube and aspirate the liquid with a pipette. 4.Washing: Add 1ml of sterile water and wash the seeds by inverting the tube for 1min, settle the seeds at the bottom of the tube and discard the liquid. Repeat this step 3-5 times. 5.Planting the seeds: Add an appropriate amount of sterile water to suspend the seeds, aspirate the suspended seeds and plant them on 1/2 MS Modified Plant Solid Medium plates (containing 0.8% agar), with 50-100 seeds per plate. After the visible water around seeds on the plates evaporated, seal the Petri dishes with sealing film.6.Break dormancy: Place the plates at 4℃ for 3 days to break seed dormancy. 7.Plant growth: Transfer the plates to a green house or a growth chamber at 21℃ with a light intensity of 120µmol m-2s-1 (6672lx) and a photoperiod of 14h light/10h dark. Seeds will germinate after several days under such conditions