7th Jul 2020
Are you ready to take your plants to the next level? With Plant Cell Tech’s Plant Preservative Mixture (PPM™), you can.
What many people don’t realize is that PPM™ is versatile. Not only can it be used in the growth medium, but it can also be used as a sterilizer. In this article, we get back to basics and look at the best way to use PPM in your gelling agent.
Protocols and Procedures
First, we will take a look at the generic procedures. Remember that each plant species may require slight modifications. If you are unsure or unable to find the specific protocols for the plant species you want to work with, you can write to our team member, Dr. Assaf Guri at firstname.lastname@example.org.
- Here’s how you can use PPM™ to simplify your tissue culture procedures: First, you will be working with your media. Add the PPM™ in the correct dosage to the media. This should be done outside the laminar hood flow. As soon as the agar solidifies, it would be best if you covered the plates. If you are dispensing the media with a pump, it would be wise to pass autoclaved hot water through the hoses before and after distributing the media.
- Most media used in the tissue culture process will be either sensitive to heat or heat stable. However, if you add PPM™ to this media, you will not have to further autoclave or sterilize it as long as you store the media in sterile containers. This ensures the stock solutions are uncontaminated and remain uncontaminated. On the other hand, if you are using rich media containing more than 200mg/l of amino acids or proteins, you should filter the media even if it contains the PPM™.
- When you are working with the laminar flow hood, some tools, like forceps and scalpels, do not need to be flamed. They should, however, be routinely dipped in 70% alcohol. The laminar flow hood is not a 100% necessity, as long as your working surfaces are kept clean and sterilized, and the work does not exceed 60 minutes.
- Naturally, the efficiency of PPM™ decreases when it is exposed to high amounts of fungal and bacterial contamination. If you are using PPM™ on in vitro germination, you should routinely sterilize your seed shells (where fungal spores commonly develop) with EPA registered bleach. Once you are ready to use PPM™, you can simply rinse the seeds with water and dry them under your laminar flow hood. If, however, your utensils or tools have been contaminated (either by being exposed to bacteria, fungi, or simply touching unsterilized areas), then you should sterilize the utensils.
- We can’t do an article about how to use Plant Cell Tech’s PPM™ without touching on the subject of dosage. The general recommended dosage for PPM™ is a range between 0.05 and 0.2%. It is essential to know that in the case of endogenous contamination, this dosage will require adjustments. In the case of high levels of endogenous contamination, you will need to rely on a higher dose to deal with the high density of endogenous contamination effectively.
When you are using PPM™ for the first transfer, it is best to insert the explants entirely into a semi-solid medium.
Plant Cell Tech’s PPM™ will make your life easier if you are working with plant tissue culture. Plant preservative mixture helps to support lab productivity, but it can be used as a sterilizer for explants. So, PPM™ can be used as a handy sterilizer for explants, a supporter of healthy root and shoot development, and it can target particular contaminations that antibiotics are ineffective against. Does this sound too good to be true? Well, if you are waiting for the catch, then here it is; PPM™ is only effective when used correctly. That means that the onus is on you, the user, to ensure that you are using the product as recommended. If you do, you have the following bonuses to look forward to:
- Eradicate waterborne contamination, airborne contamination, and even contamination that has resulted from human contact.
- Eliminate endogenous contamination from explants.
- Even at the recommended dosage between 0.5 and 2ml/l, the in vitro seed germination is not negatively impacted. The seed germination, callus proliferation, callus regeneration, and bud induction are not impaired at all by PPM™ if used at the recommended dosage.
Take a look at this protocol and procedure page if you want more details on how to use and dispose Plant Cell Tech’s PPM™ correctly.
Let us know how you used PPM™ in the comments below, and we love hearing from our fellow plant-loving friends.
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