Step 4: Sterlize the Growth Medium

For sterilization we used an old high pressure pan.

- First, pour 500 mls water, preferably destilled, into the pan
- Next, place the closed jars with medium into the pan. CAUTION: do NOT close the lid of the jar tightly! Otherwise the jars may explode while cooking!
- Additionally: put a piece of sterilization indicator tape on the lid.
- Steam the jars for 20-25 minutes into the high pressure pan.
<p>However even M(EDTA)-1(aq) would enter the bacteria cell like a negative charge salt Cl-(aq). So some Metal EDTA-1(aq) would enter the cell and damage the DNA and the cell wall structure (my Theory).</p>
<p>Fe+2 + H3EDTA &lt;&lt; &gt;&gt; Fe(EDTA H)-1(aq).</p><p>The Fe(EDTA H)-1 when exposed to the cell wall of a gram positive bacteria would REPEL due to the negative charges of both the cell wall of a gram positive and the EDTA H Fe molecule.</p>
<p>However cobalt, manganese and iron if an<strong> oxidizer</strong> like H202 is added for pH 7.0 to 7.8 would act like Co3+, Mn3+ and Fe3+.</p>
<p>All metals I work with expect aluminum would have a +2 charge. They include iron, copper, manganese, magnesium, calcium, zinc, etc. EDTA at ph 6.2 to 10.3 would produce Al(EDTA) with no charge since Al+3 would react with the oxygen bonds of the three EDTAs. Pratically inert in terms of reaction.</p><p>Al3+ (aq) + H3EDTA-3(aq) &gt;&gt;&gt; Al(EDTA). most likely a colliod or something.</p>
<p>Here is a picture of EDTA charged with a Metal with +2 state. Most of the metals I work with expect Aluminum have a +2 charge.</p>
<p>I was wrong about the Fe.</p><p>Here is the equation</p><p>Fe+2 + H3EDTA &lt;&lt; &gt;&gt; Fe(EDTA H)-1(aq).</p><p>At pH 6.0 to 10.5 the H3 EDTA dominates and forms a weird complex.</p><p>Picture below.</p><p>Here is the reference too.</p><p><a href="https://www.researchgate.net/post/Can_someone_explain_why_EDTA_needs_basic_condition_for_dissolving" rel="nofollow">https://www.researchgate.net/post/Can_someone_expl...</a></p>
<p>Here is a theory on how EDTA works with Iron.</p><p> Fe+2 (aq) +H4EDTA(aq) &lt;&lt; &gt;&gt; Fe (EDTA)-2(aq)<br>+ 2H+(aq).</p><p>This is an equilibrium chemical one. When acid is added the equilibrium shifts left producing more Fe+2 ions which if they are inside the cell produces H202. The Hydrogen peroxide (H2O2) damages cell structures, etc including DNA.</p>
<p>I plan to work with sodium EDTA and the following salts.</p><p>Iron sulfate</p><p>Copper sulfate</p><p>Magnesium sulfate </p><p>Zinc sulfate</p><p>Aluminum sulfate </p><p>Manganese sulfate</p><p>Calcium chloride</p><p><em><strong>Warning: Manganese salts are considered potent neurotoxins if inhaled, swallowed or absorbed through the skin, Always wear PPE plus a gas mask to avoid inhaling the dust. Always wear gloves (Nitrile) and goggles or face shield before using this chemical to avoid toxicity to the brain, kidneys and liver.</strong></em></p>
<p>I am continuing a project with EDTA of iron, copper, manganese, magnesium, calcium, zinc, etc. I am testing bacteria growth with fructose sugars.EDTA salt and the compound require backwashing of a filter etc (0.1 micron) recording Time (Hours) vs pH.</p>
I have pulled yeast from beer multiple times. Your best bet though is to make a yeast starter first, than pull from the starter to the growth plates. Of course you have to use beers that have been bottle conditioned( bottle with yeast to carbonate it.) <br><br>Lots of tutorials from us beer makers if you look up making beer/yeast starters.
how about SBA? ;) Just kidding. thanks for the tutorial and I am planning on saving this for my future kids!!! First to try this with my 9 y.o. sister <3
Very cool...will help me to be more cultured...haha get it...culture.
i've been looking for a way to make petri dish medium at home, this process seems a lot easier than buying the actual agar and cooking it up. what i want to do is keep a running colony of penicillium. my hypothesis is that keeping a running colony will allow the penicillin made from the mold to be effective against newer strains of bacteria like staphylococcus. <br> <br>i don't have a pressure cooker though, would it be possible to sterilize the jars in a regular boiling pot of water over the stove? <br> <br>i also have a propane stove at my house so it seems like i could inoculate the dishes there in the presence of the flame however i was wondering what being in the presence of a flame actually does? does it just move the air away from where you're working or does it actually sterilize the air from the heat? <br> <br>finally your method for getting yeast from the beer won't work because the alcohol in the beer has already killed off the yeast. it's shed it's mortal coil and now it's body adds a grainy flavor to the beer that it made.
Hi Waldosan, <br> <br>I've a protocol for the isolation of Streptomyces from mud. Streptomyces is known to synthesis antibiotics as well. I tried it out last year with students, and it worked out fine. I'll try to post it later (busy period right now: exams). <br> <br>If you don't have a pressure cooker, you can try the process called tyndallization. http://en.wikipedia.org/wiki/Tyndallization. I've never tried it, but it would be great to know your experience if you decide to try it out! <br> <br>The flame is necessary to create a sterile environment. This will prevent contamination with other micro-organisms. <br> <br>I've already isolated yeast from Westmalle, but by using YPD plates. Though it didn't always work. Most of the yeast in the beer is indeed dead.
tyndallization seems like the best way to go for that thankyou, I'm guessing it could be done in a big pot, those big ones that can make a noodle salad for an army. <br> <br>exams, i don't miss those i won't lie... <br> <br>do we know if the same people who are allergic to penicillin are also allergic to the streptomyces antibiotic? or are there common cases of being allergic to one and the other? <br> <br>this makes me want to get some of those immortal cells that they always use instead of human testing...

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