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Offlinejamesbrown
The Learner


Registered: 07/24/12
Posts: 110
Last seen: 11 years, 2 months
First MS to agar
    #17327496 - 12/03/12 03:55 PM (12 years, 1 month ago)

So I have two moldy spots on my agar with a small ring of light white fuzzyness around the.
I thought I placed one drop of MS on my agar it must have been two though because there are the two spots.
I know that MS to agar usually results in contams but I am wondering if I got some rhiz growth that I could transfer out.
In addition I could not see any spores in my syringe I ordered from a trusted seller, now this is my second time using their product and my first attempt was successful. But in the last syringe I swear that I could see the spores. 
http://i.imgur.com/5PWBi.jpg
http://i.imgur.com/m8wJu.jpg

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Offlinejamesbrown
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Registered: 07/24/12
Posts: 110
Last seen: 11 years, 2 months
Re: First MS to agar [Re: jamesbrown]
    #17327534 - 12/03/12 04:01 PM (12 years, 1 month ago)

I tried to take better pictures but these were best I could get

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Offlinetwistedty
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Re: First MS to agar [Re: jamesbrown]
    #17327545 - 12/03/12 04:03 PM (12 years, 1 month ago)

need to work on your sterile technique.

lil more practice

those would be hard to transfer from IMO

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Offlinejamesbrown
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Posts: 110
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Re: First MS to agar [Re: twistedty]
    #17327561 - 12/03/12 04:06 PM (12 years, 1 month ago)

Thanks for the advice!

Couldnt my sterile tek be perfect and my MS have some nasties in it?

Also my petris are not divided into 3 sections, could I potentially just try again on the other side of the petri?

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Offlinetwistedty
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Re: First MS to agar [Re: jamesbrown]
    #17327572 - 12/03/12 04:07 PM (12 years, 1 month ago)

id start with new batch, and i guess a bad syringe could definetly be the prob.

you could also get innoc loop and smear spores from print directly onto agar

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Offlinejamesbrown
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Re: First MS to agar [Re: twistedty]
    #17327593 - 12/03/12 04:11 PM (12 years, 1 month ago)

ya, my only problem is that I only have my spore syringes. I wanted to start agar now and not have to wait to do some BRF to either get prints or a culture tissue.

In addition the agar plates are pre sterilized from a trusted seller.

I appreciate your feedback.

But is there a possibility I could get my transfer on and either get a clean or less contamed sample from the one I have?

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OfflinePussyFart
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Re: First MS to agar [Re: jamesbrown]
    #17327598 - 12/03/12 04:12 PM (12 years, 1 month ago)

the one you have looks like mold to me....:shrug:

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Offlinetwistedty
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Re: First MS to agar [Re: jamesbrown]
    #17327604 - 12/03/12 04:13 PM (12 years, 1 month ago)

i dont know if you can salvage any of that but you can try

making your own dishes is cheaper in long run.

bout 60bux for case of 500 dishes, and then PDA from mycosupply bout another 30bux

and the PDA is just add water

couldnt be any easier

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Offlinejamesbrown
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Re: First MS to agar [Re: PussyFart]
    #17327608 - 12/03/12 04:13 PM (12 years, 1 month ago)

womp womp. ya its pretty moldy i was just hopping that ring around it was some viable rhiz growth

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OfflinePussyFart
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Re: First MS to agar [Re: jamesbrown]
    #17327621 - 12/03/12 04:15 PM (12 years, 1 month ago)

I say mix your own and use half the nutrients...that way the rhizomorphic mycellium might have a chance at out running the contams in search of food.

But that's just me.

Edited by PussyFart (12/03/12 04:15 PM)

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Offlinejamesbrown
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Re: First MS to agar [Re: PussyFart]
    #17327643 - 12/03/12 04:19 PM (12 years, 1 month ago)

YES! I remember RR saying that.

I am also not using a flow hood and am not sure at what point in this hobby I will be attempting to make one.

I know a lot of people say a lot of things about agar and still air boxes but I really want to try and have a decent success rate. I mean If people can do G2G in them  why cant I do some agar.

I dont mean to start a pro con battle about flow hoods and still air boxes, but I just want to hear some opinions directed at me.

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OfflinePussyFart
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Re: First MS to agar [Re: jamesbrown]
    #17327721 - 12/03/12 04:33 PM (12 years, 1 month ago)

Quote:

jamesbrown said:
YES! I remember RR saying that.

I am also not using a flow hood and am not sure at what point in this hobby I will be attempting to make one.

I know a lot of people say a lot of things about agar and still air boxes but I really want to try and have a decent success rate. I mean If people can do G2G in them  why cant I do some agar.

I dont mean to start a pro con battle about flow hoods and still air boxes, but I just want to hear some opinions directed at me.



I do not have a flow hood...all my agar work gets done in a still air box with close to 100% success...if the air is still, a contam blowing on your work is impossible.

Edited by PussyFart (12/03/12 04:34 PM)

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Offlinejamesbrown
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Registered: 07/24/12
Posts: 110
Last seen: 11 years, 2 months
Re: First MS to agar [Re: PussyFart]
    #17327806 - 12/03/12 04:47 PM (12 years, 1 month ago)

How much space do you have to work with in your box. Also how fo you flame sterilize, in or out the box?

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OfflinePussyFart
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Re: First MS to agar [Re: jamesbrown]
    #17327920 - 12/03/12 05:06 PM (12 years, 1 month ago)

Never flame inside a box that is filled with alcohol fumes, or things that have been wiped with alcohol, eyebrows have been lost...always flame outside the box and bring the tool in to the SAB...This is one of the many reasons a still air box is better than a glove box for this hobby.

I use a 66 qt sterlite tub for my SAB...its kinda small for G2G transfers, but for agar work it's perfect.

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Offlinejamesbrown
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Registered: 07/24/12
Posts: 110
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Re: First MS to agar [Re: PussyFart]
    #17328212 - 12/03/12 05:56 PM (12 years, 1 month ago)

Awesome info! So I should ripout my tyvek sleeves.

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OfflinePussyFart
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Re: First MS to agar [Re: jamesbrown]
    #17328489 - 12/03/12 06:48 PM (12 years, 1 month ago)

No, tyvek sleeves are fine...attached gloves are not.

Keep your sleeves and wear gloves on your hands.

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Offlinejamesbrown
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Registered: 07/24/12
Posts: 110
Last seen: 11 years, 2 months
Re: First MS to agar [Re: PussyFart]
    #17328602 - 12/03/12 07:08 PM (12 years, 1 month ago)

Getting my syringe thru the tyvek is  sleeve, post flame sterilization, is difficult. Sugjestions?

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InvisibleSpitballJedi
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Registered: 10/13/12
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Re: First MS to agar [Re: jamesbrown]
    #17328759 - 12/03/12 07:31 PM (12 years, 1 month ago)

Quote:

jamesbrown said:
Getting my syringe thru the tyvek is  sleeve, post flame sterilization, is difficult. Sugjestions?




I'm about to start my first agar this week. I was wondering the same thing.

I have tyvek sleeves tapped to my SAB. the wrists have a big rubber-band, so they're still kinda open, until i put my gloved hand through.

I thought about wearing the tyvek sleeves instead of having them attached to the box. Some people just wear gloves and skip the tyvek all together. as long as the air is still, should be fine. moveing in and out too fast prolly stirs things up a bit. I don't know, i'm just thinking.

I think i'll just practice a bunch of times and a good idea will prolly hit me. The proof is in the pudding. I'll find out soon enough if my technique is good.


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Offlinejamesbrown
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Registered: 07/24/12
Posts: 110
Last seen: 11 years, 2 months
Re: First MS to agar [Re: SpitballJedi]
    #17328817 - 12/03/12 07:40 PM (12 years, 1 month ago)

Perhaps after entering with the sleeves tapped to the box you can inverted the sleeves flame sterilize then re enter while sleeves are comming out at you. But I would like to hear what people actually do.

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InvisibleSpitballJedi
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Re: First MS to agar [Re: jamesbrown]
    #17328933 - 12/03/12 07:58 PM (12 years, 1 month ago)

the more i do this hobby, the more i realize it's not rocket science.

When I 1st started, i made things way over complicated, and made a bigger deal out things than need be. The simpler i make things, the more success i'm starting to have.

I think i'll start with no sleeves, new rubber gloves, closed door, ac off, and an ass load of spray disinfectant in the room. I'll try and pay close attention to my technique(not disturbing the air) and see how far i get.

I would like to here from others as well.


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A little civility goes a long way

The Noob Forum
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