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Offlinetrubblesome
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Registered: 11/09/19
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Re: Injecting sterile water? [Re: LadysKnight]
    #26446070 - 01/22/20 10:35 AM (4 years, 7 days ago)

Quote:

LadysKnight said:
Here are some pre-made agar plates someone recently bought:


https://www.shroomery.org/forums/showflat.php/Number/26439961#26439961




those plates

:grossman:


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OfflineDNA_Trip
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Re: Injecting sterile water? [Re: A.k.a]
    #26446165 - 01/22/20 11:24 AM (4 years, 7 days ago)

Honestly I never knew that inoculating grains by spore syringe was looked down upon and I’m being told it sucks. My last couple grows I inoculated with a spore syringe within a SAB and also open air, both, and everything worked great so I just kept doing what always worked for me.


Edited by DNA_Trip (01/22/20 11:24 AM)


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OfflineA.k.aM
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Re: Injecting sterile water? [Re: DNA_Trip]
    #26446168 - 01/22/20 11:28 AM (4 years, 7 days ago)

Yeah same with me, all my grows have been syringe to wbs with no SAB.

Idk if I got particularly clean syringes or what but they’ve been fine. I made lc with them months ago and forgot about it until recently, tested two and both look clean after two weeks.


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LAGM2020


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OfflineDNA_Trip
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Re: Injecting sterile water? [Re: A.k.a]
    #26446191 - 01/22/20 11:42 AM (4 years, 7 days ago)

The grow that you posted me that you said was from bad spawn, how long did that shoebox take to colonize and pin? Did it go a lot slower than usual?


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OfflineA.k.aM
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Re: Injecting sterile water? [Re: DNA_Trip]
    #26446208 - 01/22/20 11:52 AM (4 years, 7 days ago)

No it was normal


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LAGM2020


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OfflineDNA_Trip
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Re: Injecting sterile water? [Re: A.k.a]
    #26446217 - 01/22/20 11:56 AM (4 years, 7 days ago)

Well damn, I think I might just throw away these shoeboxes then. There's a few random white little puffs poking through, but that's about it. It definitely is going way slower, I can't tell if it's even spreading and colonizing at all like how you would regularly. What's your set up and technique whenever you do open air?


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OfflineA.k.aM
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Re: Injecting sterile water? [Re: DNA_Trip]
    #26446227 - 01/22/20 12:01 PM (4 years, 7 days ago)

Well I guess technically it wasn’t completely open, I had a cardboard box I set on it’s side and knocked up the jars inside.

But I just wiped down the lid and needle and stuck it in the jar. I had a tyvek layer with silicon ship on top and then put a piece of Micropore tape over the ship just in case.

I still incubate my jars in the same box


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LAGM2020


Edited by A.k.a (01/22/20 12:02 PM)


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OfflineDNA_Trip
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Re: Injecting sterile water? [Re: A.k.a]
    #26446334 - 01/22/20 12:52 PM (4 years, 7 days ago)

How do you do your agar? What was the easiest way you found to do it as a beginner?


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OfflinecronicrFacebook
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Re: Injecting sterile water? [Re: DNA_Trip]
    #26446377 - 01/22/20 01:14 PM (4 years, 7 days ago)

I use half pint jars. Mix agar and brf ( 10 grams agar 8 grams brf) then add 500 mls of cool water, mix it up and heat it up on the stove, once it's thick I pour it in the bottom of the jars and sterilize. The lids are just a hole covered with 2 layers of micropore tape


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It doesn't matter what i think of you...all that matters is clean spawn

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OfflineA.k.aM
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Re: Injecting sterile water? [Re: cronicr]
    #26446397 - 01/22/20 01:28 PM (4 years, 7 days ago)

Quote:

cronicr said:
I use half pint jars. Mix agar and brf ( 10 grams agar 8 grams brf) then add 500 mls of cool water, mix it up and heat it up on the stove, once it's thick I pour it in the bottom of the jars and sterilize. The lids are just a hole covered with 2 layers of micropore tape




This sounds like the best way to try it out cheap.

I bought a nice media bottle, 40 plates, a scalpel, and a small bag of premixed lme agar.

Agar really is super easy, especially pouring the plates I’ve never had a contamination in prob 140 of them. Using them without getting contams might have a little learning curve but nothing serious.


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LAGM2020


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OfflinePsychgro
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Re: Injecting sterile water? [Re: A.k.a]
    #26447763 - 01/23/20 07:42 AM (4 years, 6 days ago)

Nothing to be afraid of with agar DNA. I’m a total noob and although it hasn’t been all successful, starting out with agar has been worth it for me. I used this wood burning tool, you can pick up at harbor freight or on amazon, to cut the holes in my SAB (105qt, go big).

https://www.amazon.com/gp/slredirect/picassoRedirect.html/ref=pa_sp_atf_aps_sr_pg1_3?ie=UTF8&adId=A040549527TVYHYIYV69Y&url=%2FWalnut-Hollow-Introduction-Intermediate-Woodburners%2Fdp%2FB000VRTG8Y%2Fref%3Dsr_1_3_sspa%3Fcrid%3DI2VOB1R9UB2X%26keywords%3Dwood%2Bburning%2Btool%26qid%3D1579789241%26sprefix%3DWood%2Bbur%252Caps%252C140%26sr%3D8-3-spons%26psc%3D1&qualifier=1579789241&id=2349388636724237&widgetName=sp_atf

I bought a sleeve of petris for $12 (which I haven’t used) and have been using zip loc minis with twist caps that I picked up at Walmart. My media bottle is a glass braggs apple cider vinegar bottle (almost 1L, 976 to be exact, they sell smaller ones too).

I’ve had some contam in very few plates, I have many second and third transfers, but mostly I have a lot of dead cultures probably from spraying too much ISO or bleach solution...go for it!


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Looking to learn all about it


Edited by Psychgro (01/23/20 08:14 AM)


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OfflineA.k.aM
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Re: Injecting sterile water? [Re: Psychgro]
    #26447784 - 01/23/20 08:08 AM (4 years, 6 days ago)

Why were you using iso and bleach with agar??


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OfflinePsychgro
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Re: Injecting sterile water? [Re: A.k.a]
    #26447790 - 01/23/20 08:18 AM (4 years, 6 days ago)

To clean stuff inside the SAB prior to actually introducing the cultures...I would wipe down the sab, drench the towel on the bottom, wipe down the outside of all plates and syringes (excessively). I was too worried and scared.

Now I know it’s okay to get some contaminated plates,it doesn’t have to be perfect and can be counter productive when one tries to be.  so long as I get some good plates/transfers while I’m at it, it’s worth it. My SAB technique still sucks, I fumble around a lot but finesse will come with time


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OfflineA.k.aM
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Re: Injecting sterile water? [Re: Psychgro]
    #26447803 - 01/23/20 08:39 AM (4 years, 6 days ago)

Oh ok I thought you were putting it on the plates or something.

Man if you used enough to damage the cultures just from fumes I’m surprised you didn’t blow up at some point lol.


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OfflinePsychgro
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Re: Injecting sterile water? [Re: A.k.a]
    #26448109 - 01/23/20 12:23 PM (4 years, 6 days ago)

Quote:

A.k.a said:
Oh ok I thought you were putting it on the plates or something.

Man if you used enough to damage the cultures just from fumes I’m surprised you didn’t blow up at some point lol.





my guess is that some of it must’ve gotten on the plates from the air...only way all those plates (between 30-40) have remained “sterile” for 3 weeks 🤪.im about to toss those(20 glass petris 60mm, smaller than I like...I’ve learned now... and like 20 Ziploc minis)

The other big mistake I made was forgetting to microwave agar after adding all ingredients...I would think to myself “why isn’t everything gelling?” And I would then add water, nutes and/or agar (without keeping track of ratios. I think that and “over sanitizing” have been my biggest mistakes.

After watching the channel “home mycology” on YouTube I realized I was being too OCD with trying to sterilize things. I wouldn’t recommend what that guy does but I think a happy medium between that and what I used to do is appropriate.

DNA, go for it!! 😝


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