How to Prepare Potato Dextrose Agar (PDA) using Fresh Potatoes (Part 1/4)
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- Опубліковано 9 сер 2020
- This video will demonstrate how to prepare potato dextrose agar (PDA) using fresh potato. PDA is a microbiological agar used to isolate or culture wide varieties of fungi. With Malay subtitle.
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Eindelijk een keer een goede informatie over het maken van agar agar met aardappel bloem, dat je zelf kan maken. Het is gewoon nergens te vinden die geen andere ingrediënten zijn toegevoegd zoals zout bijvoorbeeld
Thank you for this. Very well explained
Very nicely explanation is done. Thank you !!
Nice 😍😍
Can I use glucose powder for pastry instead of dextrose, if yes, should it be the same amount?
Thanks for your help, continue teaching like this.
Great. Thanks.
Thank you from sri lanka 👌
Is that a glass or plstic growing kit
Thank you for the well-explained video. May I ask if I can use Dextrose monohydrate (which is used for animals)?
yes, it can be used as well.
very nice thank you
Thank you 👍
👍👍👍
Thank you for the detailed information🤗 very clear explanation. May I know if I can use liquid of « sabouraud dextrose » instead of dextrose powder?
yes. it can be used as well.
Hi, may i know the agar weight ratio if used with strand agar?
I'm not clear to your question. Do you mean the ratio of fresh potato to agar?
Please i want a result and descussion for my rapport
Thanks for the video! Can i use pressure cooker instead of autocleve?
yes. cook in a pressure cooker for 30-40 minutes.
How long does the PDA last? I mean, how long can it be stored and still good to be use? And what are the conditions for storing the PDA?
storage depends on the storage temperature and how u keep them... in a sealed bag, etc. if kept at room temperature, I usually recommend not more than a month. If in a chiller, it can be stored longer, >6 months (but seal to avoid loss of moisture).
is it okay to use red agar for oyster mushroom.
the colour of agar has no issue as long as it contain agar only
So do I refrigerate the agar after? Do I want it refrigerated after I colonize my plates with mycelium? How do I get best growth of mycelium? And how do I slow down growth if I want to keep it alive but not growing exponentially? Thanks!
The optimum temperature is depending on the mushroom species. For Pleurotus, they grow best within 25-28 degree Celsius. To keep them, it's best to store in 5-8 degree Celsius.
Hi sir i'm a newby volvariella volvacea cultivator, i just want to ask if there is a way or something to add in pda to make it strong enough to fight contamination?, thank you and more power.
yo may add antibiotics to suppress bacteria.
Hi, I will only use a kitchen grade pressure cooker to autoclave.. how will I know and adjust the right temperature? Thanks
just cook for 30-40 minutes, and it will be fine.
why do it need to heat the flask mouth first before pouring the media?
to destroy any possible contamination on the flask mouth
So Clean And Beautiful
Thank you! Cheers!
@@ropenedu dude what's your take on "potato flakes" and how good or bad they are for mycelium? every time i prepare with potato flakes media has pungent smell mycelium grow but takes forever. first time preparing with fresh potatoes and distilled water after watching your video, thanx
@@FrozenFlames243 Thanks for your comment. I never use potato flakes, but theoretically it should work as long as it contain pure potato flakes (no other ingredient). Potato flakes is a dehydrated from of potato which I recommend you use 39g/L. Please share if the method you tried based on this video is a success.
@@ropenedu thank you for reply, and i made 12 60mm petri dishes using the method in video two things about this method are amazing, one the petri dish is crystal clear something that does not happen with potato flakes, two mushroom cultures are loving it, its not too much nutrient and not to less just prefect thank you for this wonderful video
@@FrozenFlames243 glad to hear that. thanks.
Can you use sucrose or glucose instead of dextrose
sucrose is table sugar, and it is a disaccharide. Whereas, glucose and dextrose (both are same thing) is a monosaccharide which is easier to be taken by fungi. Therefore, use glucose (also known as dextrose).
instead autoclave.. can i use scalding technique? i dont have any tools. or should i buy pressure cooker for this step too?
I refer the term "scalding".. I quote "Scalding is accomplished in water heated to around 185 °F (85 °C), usually in a double boiler, which conducts the heat of the water, contained in a bigger pan, to a smaller pan containing the food, thus avoiding contact between food and water."
I don't think the scalding technique can work. Other than autoclave, you can steam using a steamer for 6-8 hours. A pressure cooker is better which work similarly as an autoclave.
@@ropenedu okay thanks
thanks!! In the end of the video, you keep the petri open for 5 minutes. What is the risk to maintain open like that? In relation to other fungus enter in the petri
Thanks for your comment. The media was poured in a laminar flow, with the air blower turned on, and the laminar flow is also equipped with high-efficiency particulate air (HEPA) filter that can filter out any contamination from the air. In a well-functioning laminar flow, there's very small risk of contamination on the opened petri dish. The reason of letting the lid open is to release any excess water vapour, and avoid water condensation in the petri dish. However, this method is not advised if no laminar flow facility available, where the lid must be closed as soon as possible after pouring.
@@ropenedu how if I close with paper or fabric if I don't have laminar flow??
@@muhamadaiman8253 please refer to other youtube videos where u can use a plastic container ua-cam.com/video/svUq2OeQNC4/v-deo.html
What can I use to do it for a pure liquid Mushrooms culture
you can use the broth one. not with agar
In place of the autoclave, can someone use pressure cooker (pot)?
yes. you can use a pressure cooker.
Thank you explaining it in detail. May I ask if there are other alternatives to an autoclave?
use pressure cooker.
Hello I require information. May I know why the PDA concentration was chosen at said level? I need to cite this in my lab. Thank you.
it was adapted form the ingredients in a commercial PDA powder
Thank you …❤😊
Thank you both for video. Should the any antibiotics/ fungicides added to the media?
Antibiotics can be added to prevent any growth of bacteria. Fungicide is not recommended as it will affect the growth of the mycelia of mushrooms.
Good day, Sir. The dextrose you used is it SDA or normal dextrose sugar?
Sir I am from the India and I want need to your help
Thanks
Hello, I hope you will answer my question.
Can nutritional yeast be used together with glucose for fungi growth?
Another question is why can't fungi grow alone on glucose + water ?
Is malt extract the same as simple glucose?
Can nutritional yeast be used together with glucose for fungi growth?
Yes. As long as the nutritional yeast is sterilized. The yeast can provide additional nutrients for fungal growth.
Another question is why can't fungi grow alone on glucose + water ?
Fungi can even grow on water agar. They can grow, but with poor growth as compared to addition of potato or malt extract
Is malt extract the same as simple glucose?
No. malt is polysaccharides (complex sugar). Malt extract act similarly as potato extract
@@ropenedu that was a fast reply 😍.
You're a rare gem. I wish I found your channel a while ago when I kept failing with my mycelium growth experiments. After a few years I am trying it again. Trying to learn as much as possible.
Thanks man. Your education is how god intended it
Hi, can i use edible gelatin (from animals) insted of agar?
I am not really sure, but as I know, gelatin also can be the solidifying agent for media. You can try.
Do I boil the potatoes in low heat?
Yes, low heat will cook the potatoes longer and better. The most important things is, the potatoes need to be soften.
Bolehkah hand sanitizer digantikan sebagai medium untuk sanitize
boleh. hand sanitizer utk gantikan 70% etanol.
How long does PDA Life Expectancy? Can you suggest the way keep PDA?
Poured PDA plates can be kept not more than 2 weeks, and if kept in a chiller (4-6 degree celsius) not more than 6 months. It's based on my personal experience.
In addition, keep in air-tight container or plastic to minimize water loss from the media.
can i use potato starch
yes, sure.
can we use sugar instead of dextrose? and if yes, it will be the same amount ?
Firstly, thanks for your comment. Table sugar is sucrose, a disaccharide consists of glucose and fructose. Glucose is primarily used for respiration, and fructose for tissue formation in fungi. On the other hands, glucose or also known as dextrose or D-glucose is a monosaccharide. Obviously, sucrose and dextrose are chemically different, and metabolized differently by fungi. However, both of them has same basic function, which is to provide energy for growth and respiration. The fungi need to work harder to break sucrose to monomers before they can use them, where in this case, dextrose is the best option instead of using sucrose. In terms of quantity, I'm not really sure how much is the exact quantity of sucrose to substitute 20 g of dextrose, however, the quantity is not a critical issue as long as the food source for the fungal growth and respiration is provided in the culture media, unless the media is to be used for an experiment.
you can use the same quantity of sugar, but you will see something in loop slow growth mycelium dying out and culture will take up to 40 days ore more to take over 60mm petri dish, use dextrose its easy to find and cheap a 500g bottle will last you couple of years. also sugar experiments were done with psilocybe cubensis mycelium.
can i use saline dextrose?
do u mean the medical saline dextrose?... if yes, you cannot use that as it contain very low percentage of dextrose/glucose and also contain sodium chloride.
Hello, nice video. Can you tell me is it possibile to sterilise it in microwave? Greetings
good idea. microwave should work... you may refer to this www.cdc.gov/infectioncontrol/guidelines/disinfection/sterilization/other-methods.html#:~:text=The%20microwaves%20produced%20by%20a,organism933%2C%20935%2D937.
No, but you can only use steam
Is there any other method we can sterilize our solution other than using autoclave?
Pressure cooker
First thx for this video. Second can I use maltodextrin instead of dextrose?
I never tried maltodextrin. Theoretically, maltodextrin should work as well because dextrose is easily digestible polysaccharide or absorbed rapidly as dextrose or glucose.
@@ropenedu Thank you for the fast answer. keep up with the good work please.
Where is Anniston sir
The quantity of water that used to boi the potato depands of what!?
My suggestion, add 50% extra water. Example, if u want to prepare 1000 mL, then use 1500 mL water.
Hey bro, what kind of dextrose. I tryna buy on some laboratory lab can only find a "dextrose monohydrate 5%" its liquid and can be used for human
example of dextrose that usually found in retail store www.google.com/search?q=glukolin&rlz=1C1CHBF_enMY833MY833&source=lnms&tbm=isch&sa=X&ved=2ahUKEwjizZmyx932AhWLwzgGHY3ADY0Q_AUoAXoECAEQAw&biw=1920&bih=937&dpr=1
@@ropenedu thankyou, but can i use a kind of infuse dextrose monohydrate ? (5%)
@@ropenedu How if i used a little bit old mushroom or old mushroom to my PDA ?
@@goldenage5012 You can try experimenting using infuse dextrose monohydrate 5%. I never tried it. But theoretically, as long It contains dextrose or glucose, and no other fungi inhibitor, it may work. Can use table sugar (sucrose) as well, but sucrose is disaccharides (more complex) as compared to glucose or dextrose (monosaccharides).
@@goldenage5012 Any reason for adding old mushroom as an ingredient into the PDA?... I don't see any relevance. Using potato extract, dextrose, and agar powder are sufficient.
Thank you for this. Can I use sugar to replace dextrose?
yes. However sugar is a disaccharide, whereas dextrose or glucose is a monosaccharide. Monosaccharide is easier to be absorbed as a source of energy.
Is there a recipe for making potato dextrose broth instead of potato dextrose agar?
Yes, simply omit or exclude agar as one of the ingredients. Use potato and dextrose only.
@@ropenedu Thank you so much!
Can we use peptone along with this????
Firstly, thanks for your comment. Peptone can be added, but not really necessary in the preparation of PDA. Peptone is a source of nitrogen that can enhance microbial growth.
@@ropenedu can I get your email id ???
@@thangavelganesh5294 my email is rakibmrm@ums.edu.my
If I'm using glucose syrup instead of powder, does the weight of the ingredient stay the same? Thanks for the video!
You need to check the percentage of glucose in the syrup.. is it pure 100% glucose?... if not, u need to calculate so the concentration is 20 g/L
1ml = 1gr
What is a good substitute for dextrose in PDA agar? Molasses/honey/raw sugar, any good? And in what proportions? I can't find dextrose powder anywhere :(
glucose/dextrose is the best because they are simple sugar (monosaccharide). alternatively, table sugar (sucrose) also can be used.
@@ropenedu Is it possible to use dextrin as well?
@@adampolacek4809 I don't think so. Dextrin will become 'glue' when mixed with water.
did this but all the agar sunk to the bottom even though i mixed well. didn't get a homogene media.
1. The amount og agar powder use sufficient?
2. Mix well before pouring into a plate?
Sir how to make a pure liquid Mushrooms culture
Hai. Thanks for your comment. Unfortunately, I don't have any recorded video on that. You'll need to culture into potato dextrose broth (media without agar).
Is there no risk in leaving the petri dish open for 5 minutes after the PDA has been poured inside it? Am asking because of the present of others fungi.
the petri dish are left opened to make sure no water vapour in the plate later. If u use a laminar flow, it should be fine.
can i replace dextrose with brown sugar?
yes, but not recommended because brown sugar is a complex sugar, harder to be utilized by fungi.
Hello i want to use white sugar insted of Dg what the ratio would be
Thank you
sugar can be used but not good as dextrose. Sugar is sucrose (disaccharides) which is more complex sugar as compared to dextrose (monosaccharide)
@@ropenedu only becouse i cant find it any way at all!
But for 1Liter should i use 5grams with 15g Agar and 200g potato?
Is it correct i guess
@@hsh4870 200 g of potato, 20 g of sugar, and 20 g of agar.
Glucose that we can drink can be found in retail shops as well. Find the plain flavour.
@@ropenedu Thank you so much sir, offering your knowledge and answering it was really helpful i really appreciate it.
If it's ok can i contact you on email to ask your further questions
Thank you
@@hsh4870my plesure. rakibmrm@ums.edu.my
hi if i no have dextrose can i use lactasoy milk ?
recommended to use the most simplest form of sugar, which is dextrose/glucose.
can do this for volvariella mushroom ?
@@menghongra9720 yes. same agar media can be used for isolation of Volvariella
I had only pressure cooker. How much of time I should sterilize on gas stove
I apologise ,after commenting I saw 30-40 mins is enough but in low flame or high flame and with whistle are not.
40 minutes should be fine if using a pressure cooker.
What if the more dextrose?
not necessary to add more glucose/dextrose
Is there a difference between potato dextrose broth and potato dextrose agar?
yes, only 1 difference in terms of their ingredient. PDA has agar, whereas PDB no agar. Agar is the solidifying agent in a agar media
@@ropenedu so broth is a liquid medium while agar is solid?
@@mr.purrfect6500 yes, exactly.
WHy is my petri dish/PDA moisting even if I dried them properly
do u mean water vapour inside the petri dish?
@@ropenedu yes
@@jaypark6552 Probably any of these 2 factors:
1. molten PDA poured when they're too hot, or
2. evaporation of water from PDA. I usually let the water vapour evaporate before closing the petri dish lid. This must be done inside a laminar air flow.
Hi lecturer, May I know after I sterilise the PDA in pressure cooker for 30-40mins,
1) can I directly pour the PDA into petri dish at high temperature?
2) once PDA solidify , can I directly put in the mushroom tissue? Or it need to be cool down one day?
Thanks for your comments:
1. It is not recommended to pour at high temperatures because it will create a lot of water vapour inside the petri dish that will increase the possibility of contamination, and it's dangerous and can injure.
2. Once the PDA is solidified, u can use them immediately.
Hope this helps. Thanks for your support. Please help this channel to grow by subscription, like, and share.
Please, Dr. What type of dextrose did you used, SDA or normal dextrose sugar?
it's dextrose only. normal one
@@ropenedu thank you, Sir, for your response.
Hello, firstly want to say Great Video 🙏🏻
I wonder what should i/we use "if" i/we dont have any equipments/tools like the Laboratory have, such as "Autoclave and Laminar Flow(HEPA)"
Let's say want to try it in "DIY Home Edition"
Firstly, tanks.
You can use a DIY container to substitute laminar air flow. Try to search videos on other youtube channel on how to build it.
Few examples of inoculation box
No.1: ua-cam.com/video/svUq2OeQNC4/v-deo.html
No.2: ua-cam.com/video/4vTEzc4RNOQ/v-deo.html
No.3: ua-cam.com/video/lUQDPC9HX4Q/v-deo.html
An autoclave can be replaced with a pressure cooker, steamer, or DIY steamer.
ua-cam.com/video/mlzC3skoigA/v-deo.html
ua-cam.com/video/hZsRZh1onmo/v-deo.html
Hope this helps.
@@ropenedu a very helpful reply indeed!
Do you have a video on how already made dehydrated PDA can be used to prepare this culture?
use 39 g/L water. mix them well, and then cook in autoclave or pressure cooker.
@@ropenedu 39 g/L water mixed with what quantity of dehydrated PDA?
@@ropenedu with what quantity dehydrated PDA?
@@sonotuttotuo7782 39 g dehydrated PDA and 1 litre of water.
why did the PDA media change color
Sorry, actually the recording was done separately with different batch of PDA. In addtion, the PDA will be slightly darker after sterilized or "cooked". Thanks.
what is the brand of dextrose and agar
@@ranggaadiwinata5464 in this video, I use Sigma dextrose, and QReC agar... However, the brand is not important.
are there any additional ingredients other than potatoes, agar and dextrose in the manufacture of tissue culture / PDA
@@ranggaadiwinata5464 nope, that are the 3 basic ingredients for preparation of PDA.
If this works im gonna be so happy.
Tried once with another tutorial and it failed. Nothing grew
did it work?
@@YomAlwa3eed Yeees, it did!
@@dienosorpo Thanks 👍
I use my moms pressure cooker lol
Can I make a roast in your overpriced pressure cooker?
it is an autoclave usually for lab use hehe
Your audio is so low
Voice is very low no energy...
There are people with this kind of natural voice and attitude. At least they know what they are doing. Let’s just appreciate their effort and generosity in making this video accessible to learners..
I can't get you on email anymore. Whenever I send it doesn't deliver. Please, can you help me with your email address, Dr.