Making Tissue Culture Media for 100 Deli Containers!

[@dr.greenthumbs345]

The tip about using the slow cooker liner to autoclave the deli containers is priceless. Thanks! The larger "Oven Bags" are also available if anyone is interested. Made from the same nylon blend as the slow cooker liners. They are 23" x 19" and heat resistant to 400°F / 204°C.

[@jaybien1]

YES

[@Astrid-oq3kk]

Hi, I've noticed you calibrate the PH before adding the gelling agent, why is that? Doesn't gellan gum risk impacting the PH of the medium?

[@PlantCellTechnology]

Hi Astrid, a good quality gelling agent does not change the pH of your media. The reason I calibrate the pH before adding the gelling agent is to keep the probe clean. I also added the gelling agent to the bottles.

[@Astrid-oq3kk]

@@PlantCellTechnology OK thanks :) I'm stuck with a big bag of poor quality agar at the moment (it has sugar in it, so I deduct the sugar from my recipe) so I think I'll keep measuring after adding it at the moment

[@sjoer]

You added three squirts of vitamins (one more than the others) to the left beaker!

[@PlantCellTechnology]

Great catch! It happened while making the video.

[@rafimled4868]

There is no contamination if you hold gloves over the containers??

[@PlantCellTechnology]

We need to ensure that the gloves are sterilized, clean, and have nothing stuck to them that can be shed into the culture media while using that in the process. However, we need to avoid passing our hands over cultures. We need to be as careful as we can. Though contamination in cultures depends on many factors, you need to ensure that you follow best practices to avoid it.

[@rafimled4868]

@@PlantCellTechnology In plant cultures, is it better to have horizontal or vertical filtration in a chamber?

[@rafimled4868]

@@PlantCellTechnology I bought one, I'm waiting for delivery, did I do the right thing? I also have one from you, PPM, I'm testing it BioBase BSC-1500IIA2-Pro

[@PlantCellTechnology]

@@rafimled4868 What did you buy? Laminar hood?

[@rafimled4868]

@@PlantCellTechnology Yes, (Class II A2 Biological Safety Cabinet BSC- PRO) BioBase BSC-1500IIA2-Pro (only with vertical flow and everywhere I see horizontal)

[@LilHer08]

@Plant Cell Technology - you added an extra ml of BAP to the center beaker. Time stamp 16:34. The start of you adding BAP was 16:14. Just thought I would let you know as I am not sure what the outcome could be.

[@PlantCellTechnology]

Great catch. Thanks for sharing.

[@charliemagoo7943]

in this video you seal the lids before cooling . others you seal the lids???

[@alfred282]

Great video! Thanks for sharing.. just wondering, how long do you autoclave the container and the 1 litre media?

[@PlantCellTechnology]

You need to autoclave the media at 15lbs (121°C) for 15 minutes.

[@alfred282]

@@PlantCellTechnology thanks for the reply!

[@PlantCellTechnology]

@@alfred282 Happy to help!

[@hxx117]

What is the Plant Cell Technology catalog equivalent of those extra vitamins 16:46 you applied and what is the dosage for both houseplants and carnivores? Thank you for so much knowledge, Francisco!

[@adedoyinadetunji2078]

what is the size of your pressure cooker?

[@PlantCellTechnology]

It comes in different volumes. so, depending on how much media you need to sterilize you can choose either a 5L, 10L, or 20L pressure cooker for your use.

[@alexiszisakos295]

I just started with tissue culture and was wondering if your hormones are heat stable or simply do not degrade with the temperatures in the pressure cooker? Sadly, i'm not able to buy the ingredients from PCT since the shipping is more than a hundred bucks, so i hope this counts for the hormones of other companies too.

[@PlantCellTechnology]

Some plant hormones are autoclavable and some are not. Which hormones do you want to know about? Also, we have distributors worldwide who can help you save on the shipping cost. If you can let us know where you are located, we can connect you to the closest distributor to help you with your tissue culture requirements.

[@alexiszisakos295]

@@PlantCellTechnology Thank you for your answer! I'm currently using NAA, IAA, IBA, Kinetin and BAP. I'm located in south-western Germany and would love to use the products of PCT! Also, it would be great to know with which amounts i need to compensate a potential loss of phytohormones which are partly degrading under 120°C.

[@PlantCellTechnology]

Hi Alex, Here are the details of our German distributor: They will help you with all of your tissue culture requirements. Also, we couldn't get what you wanted to say with this "Also, it would be great to know with which amounts I need to compensate a potential loss of phytohormones which are partly degrading under 120°C." Can you rephrase the part so we can help you better? Thanks! @@alexiszisakos295

[@alexiszisakos295]

@@PlantCellTechnology Sure. So the datasheet of my hormones says that all of the hormones i'm using are autoclavable (marked with a CA) while some are marked with CA/F. So the hormones are autoclavable but there is some potential degradation during autoclaving which can be compensated by adding a little more. So the question is how much is basically degrading... Is it 1/4 or half of it? (I know i can simply add them by sterile filtration 😄)

[@nejc1]

@@PlantCellTechnology Can you share details of the German distributor, I'm interested in some of your products too.

[@beckylight257]

I put my deli cups in a still air box, spray them with vinegar, let that dry, then spray with peroxide, let that dry, then spray with isopropyl and let that dry. Its not 100% but its close.

[@ShubhamKumar-fh5hd]

Can I use UV light for sterilization?

[@PlantCellTechnology]

No. For effective tissue culture media and tool sterilization, the autoclave is essential.

[@AffinityPhoto]

@@PlantCellTechnologybut you are using a pressure cooker? The autoclave is 15psi steam. The bag totally blocks the autoclaving process.

[@PlantCellTechnology]

@@AffinityPhoto The bag's content still gets some moisture inside to help with sterilization. You can add some water to the bag to help with the process.

[@AffinityPhoto]

@@PlantCellTechnology the water in the bag reaches 15 psi to penetrate the porous materials or would the sealed bag rupture?

[@PlantCellTechnology]

@@AffinityPhoto no the autoclavable bag won't rupture.

[@nakamurajinusuke]

what grow light do you use for tissue culture?

[@PlantCellTechnology]

We generally use LED lights for the tissue culture plants.

[@ShubhamKumar-fh5hd]

Is there any alternative of tween 20 ?

[@PlantCellTechnology]

You can use Triton-X-100. Get more insight on this page: www.researchgate.net/post/What_can_be_used_as_a_cheaper_alternative_of_tween_20_especially_for_plants_having_more_dense_trichomes#:~:text=We%20use%2070%25%20ethanol%20for,Good%20luck.

[@OBOI323]

Few drops of peroxide per liter

[@tzi0m]

What is the size of your containers?

[@PlantCellTechnology]

These are 12 oz containers. We sell the products we use on our website!

[@OBOI323]

Ok here’s a hack go to restraint supply buy ketchup cups and lids They are gamma sterilized, so if you just open them in front of your Flow Hood, important to them and put your lids on, you’ll be fine

[@AndresAQChilo]

Muchas gracias. Excelente video!

[@PlantCellTechnology]

Thanks! Happy to know that you enjoyed the video!

[@mikearts6836]

16:40 - pretty sure you double dipped that center one. You never missed it the first time around.

[@PlantCellTechnology]

Great catch! It happened.