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InvisibleGerms
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Registered: 06/26/11
Posts: 4,607
Loc: Texas
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Re: Pastywhyte's EzLC tek. [Re: Boogieman47]
    #24437200 - 06/26/17 05:01 PM (6 years, 6 months ago)

I got a 10 pk of 60 cc syringes, 10pk of 18guage dispensing needles and 10pk of whatmans all from amazon just made up 4 jars so we'll see how this goes. Out of 100+ dishes I've only had two with satellites so I'm excited to see how I do with LC


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Invisiblemos619
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Re: Pastywhyte's EzLC tek. [Re: Germs]
    #24439851 - 06/27/17 04:18 PM (6 years, 6 months ago)

Wanted to get clarification on a few things...this is just agar and whatever nutrient source with no honey, karo, STIR, correct?

I haven't really worked with empty syringes before and I'm concerned about possibly contaminating the inside of the barrel when flaming outside of the SAB, is that a legit concern and if so what precautions can be taken to mitigate this possibility?


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: mos619]
    #24439871 - 06/27/17 04:24 PM (6 years, 6 months ago)

PC the syringe in foil and don't unwrap until you are ready to aspirate.


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InvisibleGerms
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #24439982 - 06/27/17 04:52 PM (6 years, 6 months ago)

Quote:

Pastywhyte said:
PC the syringe in foil and don't unwrap until you are ready to aspirate.



Do you reuse syringes or get presterilized ones, and which do you prefer if youve tried both?


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OfflinePsilosopherr
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Re: Pastywhyte's EzLC tek. [Re: Germs]
    #24440934 - 06/27/17 10:11 PM (6 years, 6 months ago)

He just said PC it, indicating re-use.

I've never had a problem re-using, I'm sure they go bad eventually, hasn't happened to me.


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Invisiblemos619
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #24441561 - 06/28/17 07:51 AM (6 years, 6 months ago)

Quote:

Pastywhyte said:
PC the syringe in foil and don't unwrap until you are ready to aspirate.




Yeah I got that, but I'm just thinking...with a 60cc syringe and 120ml water, that's two aspirations. Would it be safe to just skip flaming since it's all going to the same place?


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InvisibleGerms
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Re: Pastywhyte's EzLC tek. [Re: mos619]
    #24510329 - 07/26/17 07:46 PM (6 years, 6 months ago)


This is the first stage before injecting water, I know the tek says quarter sized but I set these on a shelf and forgot about them, they're like two inches wide are they still usable? It's my last live culture of martinique so if I don't have to chunk em that'd be nice. not sure what the cons would be


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Offlinetryptkaloids
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Re: Pastywhyte's EzLC tek. [Re: Germs] * 1
    #24510347 - 07/26/17 07:53 PM (6 years, 6 months ago)

you'll be fine, I think the main reason he recommends that size it so you can still be sure it's clean while saving colonization time by getting it into the water asap


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"Remember, kids, the difference between science and screwing around is writing it down" -adam savage
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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: tryptkaloids] * 1
    #24510632 - 07/26/17 10:02 PM (6 years, 6 months ago)

Quote:

tryptkaloids said:
you'll be fine, I think the main reason he recommends that size it so you can still be sure it's clean while saving colonization time by getting it into the water asap




:whathesaid:


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OfflineManifoldPrime
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #24511036 - 07/27/17 04:24 AM (6 years, 6 months ago)

I let it get fully colonised, I want to make sure there isnt some unseen 'tam on the agar surface, any cons to this?

I finally got around to getting the 60 ml syringes, and it turns out they have these big tapering tips instead of the regular Luer slip tils. so to make them useful, I'm gonna silicone some needles INSIDE the tip and affix it, then PC them before use.

Like so, green is needle:

\|| /
  \/
  |


The other way around this would be to affix 5ml syringes on the end without the plunger as a sort of "adapter"


Again, this tek rocks my socks off. I struggle with Petri plate supply and pour tek is extra work so I've stocked up on these jars, I have like 40 125mls and 30 250mls with ship lids busy colonising as we speak :biggrin: I'll maybe take a few pics this weekend when I do my checkup

Thanks again Pasty!


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OfflineElegua
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Re: Pastywhyte's EzLC tek. [Re: ManifoldPrime]
    #24571829 - 08/22/17 01:13 PM (6 years, 5 months ago)

What do you guys think about using the silicon rings for the inside of the LC lid?  Is it useful, of no value, or can that end up as a hidden contaminant vector?

I'm asking because I've been experiencing some strange things, possibly (probably?) contaminants using this system that I don't experience with pastyplates.  My technique is the most probable problem but...just thought I'd check.


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InvisibleComebackKid
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Re: Pastywhyte's EzLC tek. [Re: Elegua]
    #24572531 - 08/22/17 06:30 PM (6 years, 5 months ago)

I can see it being useful. I've noticed plastic lids like to leak so I usually stick with metal for LC


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InvisiblePastywhyteMDiscord
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Re: Pastywhyte's EzLC tek. [Re: ComebackKid]
    #24572548 - 08/22/17 06:38 PM (6 years, 5 months ago)

I usually don't actually shake the LC, mostly just swirl it to cut down leaks. But it can be an issue sometimes. Use a gasket, pour instead of aspirate,  or use metal lids. Few ways to do it.

However if your contams are not coming from the swirling or shaking the LC then you definitely need to watch procedure. Always inspect the colony before you add water. Test it on agar if you suspect something went wrong. This requires the same care as most LC, never take liquids for granted.


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OfflineTexshrooms
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Re: Pastywhyte's EzLC tek. [Re: Pastywhyte]
    #24573426 - 08/23/17 01:10 AM (6 years, 5 months ago)

Awesome write up pasty!!! Bookmarking for future use


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OfflineElegua
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Re: Pastywhyte's EzLC tek. [Re: Texshrooms]
    #24574083 - 08/23/17 11:25 AM (6 years, 5 months ago)

Hmm, shaking may be the problem, I do tend to get a little...enthusiastic.  Hadn't thought of that, I'll work on swirling from now on, thanks!


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OfflineMcGrimm
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Re: Pastywhyte's EzLC tek. [Re: Elegua]
    #24574234 - 08/23/17 01:09 PM (6 years, 5 months ago)

bravo


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OfflineSteevo
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Re: Pastywhyte's EzLC tek. [Re: McGrimm]
    #24628273 - 09/14/17 08:22 AM (6 years, 4 months ago)

I think its time I try my hand at some LC. I've done Munch's blenderless LI a couple times but have been reluctant to venture into LC territory. No time like the present


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Invisiblestareatclouds
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Re: Pastywhyte's EzLC tek. [Re: Steevo]
    #24701172 - 10/11/17 01:09 AM (6 years, 3 months ago)

Just wanted to clarify. You're putting 60ml of sterile water and 25ml of agar media into each jar, correct?


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OfflineManifoldPrime
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Re: Pastywhyte's EzLC tek. [Re: stareatclouds]
    #24701369 - 10/11/17 05:59 AM (6 years, 3 months ago)

If a noob like me can nail down LC (with this wonderful tek of course) then you can do it . Just be vigilant and thourough with your tek and observation of growth. Be ruthless when it comes to contams.


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Please correct me if I say something wrong!

| Current Trade Print Availability:Some PESA and RW available for South African posters. Snail Mail only. |


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InvisibleSnazz
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Re: Pastywhyte's EzLC tek. [Re: ManifoldPrime]
    #24701633 - 10/11/17 08:54 AM (6 years, 3 months ago)

Definitely trying this out with my new Copelandia cultures,once my plates look nice. Heard great things about that matchup.

Made rgs soak agar plates x28 last night and have 3c of liquid in fridge.

:toast:


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