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OfflineLySergic_Dude
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Mycelium or contam
    #26611954 - 04/19/20 02:21 AM (3 years, 9 months ago)

Hey guys so I found these on my agar plates
I got a LC from a close friend so when I transferred to agar it was a bit messy the liquid did make contact with the agar but now I'm not sure if the liquid had left behind any mycelium and that's where the spread comes from or if it's a contam.

1.
2.

3.




4.


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InvisibleD3_Myc
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Re: Mycelium or contam [Re: LySergic_Dude]
    #26611980 - 04/19/20 02:50 AM (3 years, 9 months ago)

Looks bacterial to me but I’ve never dealt with LC.


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InvisibleHtaeh
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Re: Mycelium or contam [Re: D3_Myc]
    #26612012 - 04/19/20 03:34 AM (3 years, 9 months ago)

I saw someone post an example picture of bacteria on an agar plate and it looked rather similar I'm afraid, definitely wait for a trained eye! oh and get cooking some more plates in case! :wink:


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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Offlineredhandmat
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Registered: 05/09/19
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Re: Mycelium or contam [Re: Htaeh]
    #26612032 - 04/19/20 03:57 AM (3 years, 9 months ago)

Im a noob.... But it looks like its mostly bacterial with some very weak myc in there (at least its my guess). Like in the third picture,the middle very white thready half-moon shape. In my limited experience, I would take a tiny bit from it and transfer to a new and clean dish and then hot pour some agar on the old one. If you are lucky then either the new dish will show some more clean growth OR some clean growth will grow through the hot pour on the old one. In both cases, transfer the new growth asap to another new dish. And so continue until you get an entirely clean culture.

Another tip that helped this noob: with the transfers store the petri up and down! I realize this isnt possible with the original LC inoculation for obvious reasons. But now that you are working on agar, its much better.

Also, if you can do more dishes with the original LC but put as little as possible of the LC liquid on agar then do it. Im pretty confident that you will get a nice culture sooner or later.


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Offlineredhandmat
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Re: Mycelium or contam [Re: redhandmat] * 1
    #26612044 - 04/19/20 04:13 AM (3 years, 9 months ago)

ALTERNATIVELY, in the last reply I told you to try getting a single drop on the agar. BUT I went through the forums and I found a possible solution for you:
1: put a small drop on a loop (after flaming it red hot and cooling it on agar as Im sure you have seen beeing doen in many videos)

2: Streak the agar plate like this with the loop exactly like the picture and gif are showing you:



3: Store the agar plates up and down and lets see how it looks with time.

Im pretty confident this should help you get something clean that you can transfer.


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: redhandmat]
    #26612066 - 04/19/20 04:32 AM (3 years, 9 months ago)

Thank you so much guys!!!

So I've literally took that half moon looking piece lol and transferred it I'll post pics shortly

It's a tad bit cold so I'm gonna hook up my e blanket


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: LySergic_Dude]
    #26612070 - 04/19/20 04:35 AM (3 years, 9 months ago)

The inoculation loop idea is really brilliant might do that with another plate I have.

And thank odin I poured a few plates already waiting in my fridge


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: LySergic_Dude]
    #26612071 - 04/19/20 04:40 AM (3 years, 9 months ago)



I'm gonna transfer from this asap


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Offlineredhandmat
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Re: Mycelium or contam [Re: LySergic_Dude]
    #26612078 - 04/19/20 04:58 AM (3 years, 9 months ago)

Quote:

LySergic_Dude said:
Thank you so much guys!!!

So I've literally took that half moon looking piece lol and transferred it I'll post pics shortly

It's a tad bit cold so I'm gonna hook up my e blanket




Dont forget to do a hotpour as well. Like I was in the same situation you were on a few weeks ago. Only I think my myc was stronger. I did both the hot pour and transfers. And the hotpour+ place up and down really helped. I transfered the results of hot pour to a plate, let it grow and 6 days ago I put the results in grain. So far its looking really amazing. My first transfer plates did not look as good as the transfer from hot pour.

So I guess my advice to you is to try as many different things as you possibly can: and work as cleanly as you can: Shower, pc for appropriate time, clean working are and always use SAB (if you dont have flowhood), limit your hand movements inside SAB by always having a strategy on what to do before you do it, noc as many plates as you can, etc. Altogether, you will maximize your chances of getting clean growth.

And one important thing: in the beginning, see if you can blank noc a plate (pretend tot noc it but dont actually put any spores in it) just to see if it grows any contams later. That way you will know if you did everything cleanly or not.

Good luck!

PS!
If you like me don't have an actual SAB (my gf dont want to have big empty storage bins lying around), then be creative with plastic wrap and tape. For example I remove the clothes from a clothing shelf in our wardrobe and just clean it really well with soapy water (you can also wipe it down with lysol), use plastic wrap and tape to make it into a "box" while leaving some room to put in my arms. Works super nice, and I haven't gotten any contams in it.



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Offlineredhandmat
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Re: Mycelium or contam [Re: LySergic_Dude]
    #26612084 - 04/19/20 05:09 AM (3 years, 9 months ago)

Quote:

LySergic_Dude said:


I'm gonna transfer from this asap




And store up and down, I swear we underestimate the effects of storing plates properly. like having all that water rolling around the agar has not worked well for me. We could argue that everything inside the agar plate should be sterilized after 20-30 min at 15 psi.... Buuuut obviously us people using a SAB are gonna open up our plates in a non-sterile environment. Sure we are counting on phisics to help us keep the plate clean by letting the air be as still(hence Still Air Box) as possible. But Im prety use we still get some small contams in. Storing it upside down may increase the chances of these contams not moving around.


Edited by redhandmat (04/19/20 05:15 AM)


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Offlineredhandmat
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Re: Mycelium or contam [Re: redhandmat]
    #26612098 - 04/19/20 05:31 AM (3 years, 9 months ago)

Sorry for spammin your thread so much. I just wanted to add that you shouldnt have too much hope on the plates you have so far, as I looked back on my old bacterial plates with some myc on them; and the myc was much much stronger. Make more new plates with the tecs we discussed in the thread and Im sure you will get some results within a couple weeks :laugh:


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: redhandmat]
    #26612184 - 04/19/20 07:05 AM (3 years, 9 months ago)

Lol it's cool and plus I get tons of helpful information and for that I'm sincerely grateful

I'll definitely put that into practice


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: redhandmat]
    #26612189 - 04/19/20 07:08 AM (3 years, 9 months ago)

Quote:

redhandmat said:
Quote:

LySergic_Dude said:
Thank you so much guys!!!

So I've literally took that half moon looking piece lol and transferred it I'll post pics shortly

It's a tad bit cold so I'm gonna hook up my e blanket




Dont forget to do a hotpour as well. Like I was in the same situation you were on a few weeks ago. Only I think my myc was stronger. I did both the hot pour and transfers. And the hotpour+ place up and down really helped. I transfered the results of hot pour to a plate, let it grow and 6 days ago I put the results in grain. So far its looking really amazing. My first transfer plates did not look as good as the transfer from hot pour.

So I guess my advice to you is to try as many different things as you possibly can: and work as cleanly as you can: Shower, pc for appropriate time, clean working are and always use SAB (if you dont have flowhood), limit your hand movements inside SAB by always having a strategy on what to do before you do it, noc as many plates as you can, etc. Altogether, you will maximize your chances of getting clean growth.

And one important thing: in the beginning, see if you can blank noc a plate (pretend tot noc it but dont actually put any spores in it) just to see if it grows any contams later. That way you will know if you did everything cleanly or not.

Good luck!

PS!
If you like me don't have an actual SAB (my gf dont want to have big empty storage bins lying around), then be creative with plastic wrap and tape. For example I remove the clothes from a clothing shelf in our wardrobe and just clean it really well with soapy water (you can also wipe it down with lysol), use plastic wrap and tape to make it into a "box" while leaving some room to put in my arms. Works super nice, and I haven't gotten any contams in it.





Quote:

redhandmat said:
Quote:

LySergic_Dude said:
Thank you so much guys!!!

So I've literally took that half moon looking piece lol and transferred it I'll post pics shortly

It's a tad bit cold so I'm gonna hook up my e blanket




Dont forget to do a hotpour as well. Like I was in the same situation you were on a few weeks ago. Only I think my myc was stronger. I did both the hot pour and transfers. And the hotpour+ place up and down really helped. I transfered the results of hot pour to a plate, let it grow and 6 days ago I put the results in grain. So far its looking really amazing. My first transfer plates did not look as good as the transfer from hot pour.

So I guess my advice to you is to try as many different things as you possibly can: and work as cleanly as you can: Shower, pc for appropriate time, clean working are and always use SAB (if you dont have flowhood), limit your hand movements inside SAB by always having a strategy on what to do before you do it, noc as many plates as you can, etc. Altogether, you will maximize your chances of getting clean growth.

And one important thing: in the beginning, see if you can blank noc a plate (pretend tot noc it but dont actually put any spores in it) just to see if it grows any contams later. That way you will know if you did everything cleanly or not.

Good luck!

PS!
If you like me don't have an actual SAB (my gf dont want to have big empty storage bins lying around), then be creative with plastic wrap and tape. For example I remove the clothes from a clothing shelf in our wardrobe and just clean it really well with soapy water (you can also wipe it down with lysol), use plastic wrap and tape to make it into a "box" while leaving some room to put in my arms. Works super nice, and I haven't gotten any contams in it.





Omg that closet SAB is next level innovation


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InvisibleHtaeh
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Registered: 10/14/18
Posts: 320
Re: Mycelium or contam [Re: LySergic_Dude]
    #26612231 - 04/19/20 07:28 AM (3 years, 9 months ago)

Isn't it just lol, it got me contemplating to be fair hmm.. hahah.


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: Htaeh]
    #26612235 - 04/19/20 07:31 AM (3 years, 9 months ago)

Same here lol it's a pity I already constructed a SAB


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OfflineHamHead
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Re: Mycelium or contam [Re: LySergic_Dude] * 3
    #26612250 - 04/19/20 07:42 AM (3 years, 9 months ago)

IMO, all four plates are shit and need to restart.

I don't see any signs of mycelium in any of those pics. All appear to be bacterial.

I'm not experienced with LC either, only print to agar, after a single failed run of syringe to grain.



If I don't see cube mycelium that looks similar to this, I question if it's mycelium. Other active species may have mycelium that appears different, but I don't believe I've seen any that appear, er, slimy.

Sorry so break it to you.


--------------------
The Italian researchers’ findings, published by the INT’s scientific magazine Tumori Journal, show 11.6% of 959 healthy volunteers enrolled in a lung cancer screening trial between September 2019 and March 2020 had developed coronavirus antibodies well before February.

https://www.reuters.com/article/us-health-coronavirus-italy-timing-idUSKBN27V0KF

This online first version has been peer-reviewed, accepted and edited,  but not formatted and finalized with corrections from authors and proofreaders

https://www.icandecide.org/


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Offlineredhandmat
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Re: Mycelium or contam [Re: HamHead] * 2
    #26612305 - 04/19/20 08:07 AM (3 years, 9 months ago)

Quote:

LySergic_Dude said:
Same here lol it's a pity I already constructed a SAB




Hahaha, happy you guys find it cool :smile:


Quote:

HamHead said:
IMO, all four plates are shit and need to restart.

I don't see any signs of mycelium in any of those pics. All appear to be bacterial.

I'm not experienced with LC either, only print to agar, after a single failed run of syringe to grain.



If I don't see cube mycelium that looks similar to this, I question if it's mycelium. Other active species may have mycelium that appears different, but I don't believe I've seen any that appear, er, slimy.

Sorry so break it to you.




Nah, I think he knows his chances are slim on those. One of my fist and second agar attempts looked like that and mold: never becoming anything worth. Third agar go was a charm, but ofc since it was from a print so needed a few transfers etc. I think OP just need to not be discouraged and keep at it. I think youll get some growth withing 2-3 weeks if you try a bunch of plates. Provided the LC has any actual spores in it. Ofc if the friend that got you that LC could give you a shroom to clone or a bit of a print it would augment your chances.


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: redhandmat]
    #26612406 - 04/19/20 08:40 AM (3 years, 9 months ago)

Awww man bit bummed but yeah I'm gonna have to get new spores,culture or spawn soon


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InvisibleHtaeh
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Re: Mycelium or contam [Re: LySergic_Dude]
    #26612485 - 04/19/20 09:07 AM (3 years, 9 months ago)

Don't worry about it too much, it can be a trying experience getting into the swing of it etc, I only just started agar, threw five plates out in the garden the other day thinking I grew bacteria, I now think all my plates are fine haha..

Red has given brilliant suggestions for you to keep yourself entertained with in the mean time with the possibility of getting clean plates by the end of it, if you go for it keep us updated I'm interested in how it goes.


--------------------
"To write is to wage war" ~ Voltaire
"The Drug Medical System cannot bear examination. To explain it would be to destroy it.." ~ R.T. Trall M.D.
"A single lie destroys a whole reputation for integrity." ~ Baltasar Gracian (1601-1658), Spanish philosopher 
"We'll know our disinformation program is complete when everything the US public believes is false."  ~  William J. Casey, 1981
"Fear always works to influence the population, when the population is uneducated."
"The secret of freedom lies in educating people, the secret of tyranny is in keeping them ignorant." ~  Maximilien Robespierre
"Knowledge makes a man unfit to be a slave."  ~  Frederick Douglass


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OfflineLySergic_Dude
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Re: Mycelium or contam [Re: Htaeh]
    #26614293 - 04/20/20 03:51 AM (3 years, 9 months ago)

I'll definitely keep you guys posted and thanks so much for the input


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