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Offlinehamloaf
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Question: Culture Media. Peptone and Nutritional Yeast.
    #12791447 - 06/23/10 04:04 PM (13 years, 11 months ago)

Guys,


It is understood that peptone is a protein hydrosylate which enhances mycelia growth.
Peptone is used as an additive when making culture media.

It is as well understood that nutritional yeast is a source of extra nutrition and is
also an additive which enhances mycelia growth for any culture media.

How much peptone and nutritional yeast per culture media do you know to add to your
culture mediums? Do you add it by weight or by volume?


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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: hamloaf]
    #12792202 - 06/23/10 06:09 PM (13 years, 11 months ago)

I havent used yeast in a long time, I replaced it with peptone.

I used both at 0.1-0.3% w/v. You could go higher but that depends what else you wanna put in.

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Offlinehamloaf
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: badman]
    #12792848 - 06/23/10 07:48 PM (13 years, 11 months ago)

Thanks badman,


The thing is, for research light malt extract/dextrose LC recipe is used. I was looking
to spruce up the conventional light malt extract/dextrose recipe using peptone and
nutritional yeast. Have you heard of Malt yeast peptone agar? (MYPA). Research is
currently being conducted using this agar.

Peptone is said to be a culture media additive. Implying that peptone could be added to
an agar media and a liquid culture media. Nutritional Yeast is used in both agar media
and liquid culture media. If not done already. Research is looking to create a light
malt extract/dextose/yeast/peptone liquid media and was wondering how much of each should
be added. (MYPDex liquid media)

Any thoughts on this?
Would I still want to add peptone at 0.1-0.3% w/v?
Or would I want to go with more or less?

Anybody?


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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: hamloaf]
    #12793010 - 06/23/10 08:23 PM (13 years, 11 months ago)

MYPA will work easy.

For the MYPD LC Id go 1, 1, 0.1, 0.1%.

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InvisibleCheeWiz
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: badman]
    #12796590 - 06/24/10 01:01 PM (13 years, 11 months ago)

Hi; Peptones are added to a media to add amino acids. It's made by digesting a protein source in a hydrochloric acid solution of around 4.7PH and then treating that solution with peptic enzymes (the pep part of peptone). The end product is dried and ground into a fine power. The type of peptone is determined by the source of the protein and what peptic enzymes that were used in making it.

Biological yeast extract is add to a media as a way of adding vitamins (mostly Bs), but also adds some amino acids and chitin at the same time. It's made from autolyzed nutritional yeast. Again it's dried and ground to a fine powder.

A good over all media for mycology is: 20 grams light or wheat DME, 6 grams dextrose, 2 grams peptone, and 1 gram yeast extract to this many people will add .75 gram calcium sulfate and .50 grams magnesium sulfate and a gram of 5.6PH phosphate buffer power made up of potassium and dibasic potassium phosphate. To make an agar for plating just add 1.5% agar power to the base media solution.

There is a produce that can be found at many brewing supply stores that’s Wyeast yeast nutrient. http://www.midwestsupplies.com/wyeast-yeast-nutrient-1-5-oz.htmlIt's a mix of water and nutrient salts, peptones and yeast extract. I only recommend using the Wyeast brand as it finely ground and goes into solution easily.

A universal media base I make is nothing more than 20 grams wheat DME, 6 grams dextrose and 5 grams Wyeast yeast nutrient into a liter of DI water. To make an agar just add 15 grams of agar into a liter of the base or 1.5% agar into the media base solution. When preparing media you want to heat only long enough to get everything in solution. The same is true when sterilizing it. It's better to break a liter down into two 750ml liquor bottles like RR uses for 20 minutes than using one large flask for 45 minutes because heat will break the nutrients you've added down.

You should replace your media supplies every year with the exception of the agar its self that will last for many years if keep tightly sealed and stored in a cool area. So it's a one stop shopping thing for me every year to buy wheat DME. Dextrose and the Wyeast yeast nutrient on line. I hope this has been of some help.

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Offlinefungus_tao
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: CheeWiz]
    #12796791 - 06/24/10 01:36 PM (13 years, 11 months ago)

Try this recipe:

14g agar powder
14g lite malt extract
1g  nutritional yeast
1g  peptone
500 ml water

That's the same as the pre-mix I sent you. Works pretty well for me.
Tao


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Invisiblefastfred
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: fungus_tao]
    #12798971 - 06/24/10 07:46 PM (13 years, 11 months ago)

0.5-2g yeast extract and 1-10g peptone, with about 2g being average.

I don't know about the yeast nutrient that Hipster suggests.  One of the main reasons to use yeast extract is that it has a lot of enzymes that often help get fastidious organisms kickstarted.  I wouldn't expect yeast nutrient to have the same effect.


-FF


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I find the whole "my drug should be legal but yours should be illegal" mindset disgusting and hypocritical. It's what George Bush and company do when they drink a cocktail and debate the best way to imprison marijuana users. -Diploid

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Offlinefungus_tao
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: fastfred]
    #12799561 - 06/24/10 09:14 PM (13 years, 11 months ago)

Quote:

fastfred said:
0.5-2g yeast extract and 1-10g peptone, with about 2g being average.

I don't know about the yeast nutrient that Hipster suggests.  One of the main reasons to use yeast extract is that it has a lot of enzymes that often help get fastidious organisms kickstarted.  I wouldn't expect yeast nutrient to have the same effect.


-FF



If I understand that right, you're saying using the nutritional yeast for when your attempting to germinate spores, and skip it for preservation?
Tao


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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: fungus_tao]
    #12799584 - 06/24/10 09:19 PM (13 years, 11 months ago)

You can try it, but you really don't need yeast for anything.  PDA and MEA work just fine as is.  There's really no point in supplementing the media further.

I wouldn't bother with yeast except for fastidious (hard to grow) or really slow growing organisms.


-FF

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InvisibleCheeWiz
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: fastfred]
    #12800016 - 06/24/10 10:37 PM (13 years, 11 months ago)

Hi; Biological yeast extract is the same thing as the nutritional yeast that you buy at health food stores. It's a strain of yeast that's grown for the production of vitamin Bs. At the end of its growth cycle it's aloud to die and during this period of time it release enzymes that break down it cell wall, releasing its vitamins (autolyzed). It's dried and ground in to a powder that used mostly in the manufacture of vitamins. To the best of my knowledge most of the enzymes are broken down during the drying proses. As I stated earlier it is added to add vitamins to a media and that's from DIFCO handbook.

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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: fastfred]
    #12800509 - 06/25/10 12:29 AM (13 years, 11 months ago)

Quote:

fastfred said:
  One of the main reasons to use yeast extract is that it has a lot of enzymes that often help get fastidious organisms kickstarted.  I wouldn't expect yeast nutrient to have the same effect.
-FF




I just got off the phone with my son in law not to long ago about this because I've never have heard of that before and was told that's nothing but total Bull Shit.

I don't mind paying top dollar for agar-agar form Fungi Perfecti or Sporeworks as I know that it will keep if I seal it back up with parafilm after using. But I like to replace my other media making supplies yearly. The Wyeast yeast nutrient only cost me a few dollars and again it's a one stop shopping thing that I do every year, to buy 1 lb DME $4.75, Corn sugar (Dextrose) 5 oz $ .90 and Wyeast yeast nutrient $2.25 for a total of $7.90 and around $8.00 post for a grand total of 15.90. Now I don't think that's very much to spend and what I've not used just gets tossed out at the end of the year. Peptone runs about $16.00 for a
100 gram bottle plus about $8.00 for post and I've always had trouble with it caking up in a few months after being opened. Even when resealed with parafilm.

I'm not stating the Wyeast works any better but I've personally have had very good luck with it and I've been in this hobby off and on since the mid 70's!

Edited by CheeWiz (06/25/10 12:40 AM)

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OfflineRogerRabbitM
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: CheeWiz]
    #12801849 - 06/25/10 10:40 AM (13 years, 11 months ago)

Quote:

I don't mind paying top dollar for agar-agar form Fungi Perfecti




I should probably be ashamed of myself, but with all the other stuff going on around here, this is the path of least resistance.  I've been using it for a long time, and since even at $30/pound, it will last six months or more, it's a hell of a lot easier than mixing my own.  The best part is I don't have to boil it first, so it's quick.  Just mix the powder with cold water and autoclave. :wink:
RR


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OfflinePrimalSoup
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: CheeWiz]
    #12803695 - 06/25/10 04:34 PM (13 years, 11 months ago)

Quote:

There is a produce that can be found at many brewing supply stores that’s Wyeast yeast nutrient. http://www.midwestsupplies.com/wyeast-yeast-nutrient-1-5-oz.htmlIt's a mix of water and nutrient salts, peptones and yeast extract. I only recommend using the Wyeast brand as it finely ground and goes into solution easily.




Thanks for this info! :thumbup:

Peace
-PS


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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: PrimalSoup]
    #12805155 - 06/25/10 09:36 PM (13 years, 11 months ago)

Hi; RR needs nobody to amen for him as the body of his work stands on its own. There are some points that I personally disagree with him on, but that's life in the real world. When it comes to buying ready to use dry media from Fungi Perfecti or Sporeworks that's way to go for 90% of you.

It's funny people will spend $50.00 or more dollars on spores from a company like Sporeworks but will get annally tight assed about buying a good medium and a sleeve of sterile plates from the some company to grow them on. Hay man I just got these spores from Acme and can some body tell me how I can grow these thing using some burgers, last weeks oatmeal and a bag of ass scratchings that I've been saving. I don't know how RR and the other Mods hold it together; my hat is off to you!!

I come from a time when thing's were hard to find and I've had learned how to buy my media supplies from a number of different suppliers including Brewing suppliers. In fact a number of years ago you could even buy agar-agar though them as it could be used as a clarifying agent. Those days are gone and there is no reason not to buy and use a ready to use media power and in the long run it's cheaper to use. People like Paul and Workman have put their blood and sweat into testing them for us.

Edited by CheeWiz (06/27/10 12:57 AM)

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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: CheeWiz]
    #12815128 - 06/27/10 10:41 PM (13 years, 11 months ago)

Quote:

Those days are gone and there is no reason not to buy and use a ready to use media power and in the long run it's cheaper to use.




It's not cheaper in the long run.  Get your plates from a real discount supplier like cynmar and make your own media.

Freshly prepared media is superior to dried media and it's orders of magnitude cheaper.  If you want to spend a 30x markup for media that's not quite as good as what you can make for yourself then go ahead I guess.

Quote:

Biological yeast extract is the same thing as the nutritional yeast that you buy at health food stores.




You're all over the board here.  Earlier you suggested using "yeast nutrient".  "Nutritional yeast" and "yeast nutrient" are NOT the same thing.  Yeast nutrient is a mix of vitamins and trace elements for helping grow yeast.  It has nothing to do with yeast extract or nutritional yeast.

If you think yeast nutrient is a good media additive then fine, but don't go confusing people who are looking for actual yeast extract media.

Quote:

To the best of my knowledge most of the enzymes are broken down during the drying proses.

I just got off the phone with my son in law not to long ago about this because I've never have heard of that before and was told that's nothing but total Bull Shit.




Don't know why you would think that.  We're not talking about large proteins here.

Quote:

Yeast extract is a mixture of amino acids, peptides, water soluble vitamins and carbohydrates and can be used as additive for culture media.




So it certainly does provide an assortment of peptides.  In micro class we were tought that this mix of peptides, aminos, and other nutrients has many growth stimulating properties.  It would be rather foolish to assume that the component nutrients are the sole reason yeast extract is able to accelerate the growth of difficult to culture organisms.


-FF

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InvisibleCheeWiz
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Re: Question: Culture Media. Peptone and Nutritional Yeast. *DELETED* [Re: fastfred]
    #12826818 - 06/29/10 11:58 PM (13 years, 11 months ago)

Post deleted by Hipster

Reason for deletion: The post was beneath me!


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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: CheeWiz]
    #12826943 - 06/30/10 12:36 AM (13 years, 11 months ago)

Unfortunately, professionals have to deal with people they disagree with.  They don't cling to positions they can't defend, nor do they run off crying every time they lose an argument.  They don't resort to name calling or crybabyism.  They support their positions with facts and don't make accusations without supporting evidence.

RR and I were both booted from topia because Hippie3 was a dushebag.  The first time I disagreed with him and proved him wrong I was unceremoniously banned.  The things ignorant little shitstains like him hate more than anything is facts, literature, and supporting evidence.  You seem to take after him in more than name.


-FF

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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: fastfred]
    #12828164 - 06/30/10 11:11 AM (13 years, 11 months ago)

It’s not that I don't want to deal with people that I disagree with. I will leave to go back to China to work on a project there in the next week. I will be attending meeting and at times I will be the dumbest person in the room. But I doubt that I will run across a person that as egotistic and pretences as you are Fred. I dislike you because you're a BSing egomaniac that's more of a distraction than anything. You just like to hear yourself talk and no matter how many books one could sight you're going to always feel you are right when you’re not. So this is my swan song for the next few months.

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OfflineRogerRabbitM
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Re: Question: Culture Media. Peptone and Nutritional Yeast. [Re: fastfred]
    #12828330 - 06/30/10 11:43 AM (13 years, 11 months ago)

Quote:

fastfred said:

Freshly prepared media is superior to dried media and it's orders of magnitude cheaper.  If you want to spend a 30x markup for media that's not quite as good as what you can make for yourself then go ahead I guess.





You're absolutely correct.

However, when one figures labor into the mix it changes the equation.  Mrs Rabbit and I are spending 12+ hours per day doing what we already are.  In addition to making our own spawn, we're busy mixing sawdust substrates and sterilizing 100 pounds per day, and then inoculating it the next.  We have to generate 20 pounds of grain spawn per day to keep up with the sawdust, and we also have to pick and package 20lbs to 70lbs per day of finished product and get it to market.  Add to that the chore of removing spent and contaminated substrates from the fruiting area and all the spraying with water and other maintenance, and our hands are seriously full already without making our own agar media.

With all of that, spending too much on agar is a minor part of doing business.  Many mushroom farms buy their own spawn from other labs, even though it's exponentially more expensive than making it themselves.  One has to balance all the costs of production and make the best decision possible.

Let's all get along please.  Hippie3 is now deceased and 6 feet under the ground. Nobody is going to get booted from here for having a different opinion from anyone else.  All we require is respect for other members experience and ideas. We all have something to add.  Besides, nobody wins arguments on the internet.
RR


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