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OfflineRogueTrippeR
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MS Syringe Directly to Agar - how to?
    #14503609 - 05/24/11 10:14 AM (12 years, 9 months ago)

I've tried the search but did not find an answer or tek, maybe because it's so obvious.  I want to start working on isolating on agar using spores that are in a MS syringe.  I suppose I just put the smallest drop of liquid on the agar plate and wait?  I have done it with a spore print, tissue and a piece of colonized grain but have not tried applying a MS liquid solution to a plate yet.  Any suggestions/ tips?


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OfflineBiffzilla
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Re: MS Syringe Directly to Agar - how to? [Re: RogueTrippeR]
    #14503620 - 05/24/11 10:16 AM (12 years, 9 months ago)

i think a tiny drop should do shouldnt be anything out of the ordinary ... do u have a flow hood, or glove box, or what do you use a vacuum cleaners hepa ? haha


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InvisibleHipsterDoofus
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Re: MS Syringe Directly to Agar - how to? [Re: RogueTrippeR]
    #14503638 - 05/24/11 10:20 AM (12 years, 9 months ago)

#2411747 - 03/09/04 07:39 PM

Quote:

RogerRabbit said:
When using a spore syringe with agar, simply use one drop in the very center of the dish. Try to keep the dish flat when you wrap the parafilm around it to keep that drop right in the center. It will work better for you that way.



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OfflineRogueTrippeR
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Re: MS Syringe Directly to Agar - how to? [Re: Biffzilla] * 1
    #14503643 - 05/24/11 10:21 AM (12 years, 9 months ago)

Thanks.  I thought so.  Just though of the drop of water sliding around on the plate :wink:

I use a hood.

Quote:

HipsterDoofus said:
#2411747 - 03/09/04 07:39 PM

Quote:

RogerRabbit said:
When using a spore syringe with agar, simply use one drop in the very center of the dish. Try to keep the dish flat when you wrap the parafilm around it to keep that drop right in the center. It will work better for you that way.







That was what I was looking for.  thank you.  :peace:


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Edited by RogueTrippeR (05/24/11 10:23 AM)

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OfflineLord_McLovin
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Re: MS Syringe Directly to Agar - how to? [Re: RogueTrippeR]
    #14503647 - 05/24/11 10:23 AM (12 years, 9 months ago)

Where is the problem? I mean just flame sterilize your needle and put one or two drops of the MS solution onto each agar filled plate.
IMO it is a good thing to do this if you are not sure whether the syringe is 100% clean and/or want to inoculate lots of jars with just one syringe.
I actually prefer isolating from clones rather than isolating from MSC.

:2cents:


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OfflineRogueTrippeR
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Re: MS Syringe Directly to Agar - how to? [Re: Lord_McLovin]
    #14503666 - 05/24/11 10:26 AM (12 years, 9 months ago)

Quote:

Lord_McLovin said:
Where is the problem? I mean just flame sterilize your needle and put one or two drops of the MS solution onto each agar filled plate.
IMO it is a good thing to do this if you are not sure whether the syringe is 100% clean and/or want to inoculate lots of jars with just one syringe.
I actually prefer isolating from clones rather than isolating from MSC.

:2cents:



No problem.  Yes, it is simple.  I just wanted to make sure it was a safe way to go.  It seemed very straight forward but just wanted to check as most seem to not go this route.


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InvisibleSpector
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Re: MS Syringe Directly to Agar - how to? [Re: Biffzilla]
    #14503672 - 05/24/11 10:27 AM (12 years, 9 months ago)

Quote:

RogerRabbit said:
Contamination.  Any petri dish that is not wrapped in parafilm and/or touched with bare hands is going to become contaminated.

My method is to pour and work with petri dishes in front of a laminar flow hood while wearing surgical mask and gloves.  Then, wrap with parafilm, and while in front of the flow hood, each dish is placed into a ziplock bag, which is then inflated with sterile air from the flowhood. This provides a double barrier of sterile protection around the mycelium.
RR




Quote:

Doc_T said:
I use gloves and ziploc bags, but yeah, you gotta seal them somehow.






You've probably alread read these butt, here they are anyways =]
Quote:

Roadkill said:
Yes you can work with agar without using a floowhood or a glovebox.

You would have to clean the living crap out of your work area though...and make sure that you don't have any drafts of air what so ever.

Your contam rate will be higher too.

I used to do agar work without using either...but I was extremely sterile in everything that I did.

I'd suggest building a cheap glove box...it will be a really good investment.

take care!~




This is all stuff that came to mind reading your post.

Edited by Spector (05/24/11 10:29 AM)

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OfflineLord_McLovin
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Re: MS Syringe Directly to Agar - how to? [Re: RogueTrippeR]
    #14503733 - 05/24/11 10:48 AM (12 years, 9 months ago)

RogueTrippeR said:
Quote:

No problem.  Yes, it is simple.  I just wanted to make sure it was a safe way to go.  It seemed very straight forward but just wanted to check as most seem to not go this route.




The reason is that not every cultivator here is into agar - and the ones that are usually know how to do such things.
Anyway, it is indeed better to ask how something works rather than doing terrible mistakes. :smile:
You should also know that isolating strains takes LOADS of dishes and even if you got to the point of having 10 or more isolates you still don't know whether they are actually fruitable.

Edited by Lord_McLovin (05/24/11 10:49 AM)

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OfflineRogueTrippeR
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Re: MS Syringe Directly to Agar - how to? [Re: Lord_McLovin]
    #14503793 - 05/24/11 11:07 AM (12 years, 9 months ago)

Spector, thank you for all the information :smile: and looking out for me? :peace:

Quote:

Spector said:
Quote:

RogerRabbit said:
Contamination.  Any petri dish that is not wrapped in parafilm and/or touched with bare hands is going to become contaminated.

My method is to pour and work with petri dishes in front of a laminar flow hood while wearing surgical mask and gloves.  Then, wrap with parafilm, and while in front of the flow hood, each dish is placed into a ziplock bag, which is then inflated with sterile air from the flowhood. This provides a double barrier of sterile protection around the mycelium.
RR




Quote:

Doc_T said:
I use gloves and ziploc bags, but yeah, you gotta seal them somehow.






You've probably alread read these butt, here they are anyways =]
Quote:

Roadkill said:
Yes you can work with agar without using a floowhood or a glovebox.

You would have to clean the living crap out of your work area though...and make sure that you don't have any drafts of air what so ever.

Your contam rate will be higher too.

I used to do agar work without using either...but I was extremely sterile in everything that I did.

I'd suggest building a cheap glove box...it will be a really good investment.

take care!~




This is all stuff that came to mind reading your post.




Quote:

Lord_McLovin said:
RogueTrippeR said:
Quote:

No problem.  Yes, it is simple.  I just wanted to make sure it was a safe way to go.  It seemed very straight forward but just wanted to check as most seem to not go this route.




The reason is that not every cultivator here is into agar - and the ones that are usually know how to do such things.
Anyway, it is indeed better to ask how something works rather than doing terrible mistakes. :smile:
You should also know that isolating strains takes LOADS of dishes and even if you got to the point of having 10 or more isolates you still don't know whether they are actually fruitable.




Lord, true, I am moving at light speed, so in the process I seem to skip some of the basics that, eventually I need to come back to.  :smile:

I don't think it takes as many dishes as you suggest and you know how your isolates work as soon as you grow them out.  That's the point anyway right.  Thank you for you help too.  Take care. talk to you soon.

I got what I needed so I'm done here unless it is helping someone else.

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