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j3rk0ff
Stranger


Registered: 09/07/13
Posts: 80
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agar help
#18914991 - 10/01/13 08:10 AM (10 years, 3 months ago) |
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so i cooked up my agar
500ml water 10G agar 10G LME 2G DEX
45 min @ 15 psi
so now that they are cooled im wanting to start some spores to agar. my question is since im working in a S.A.B. is flame sterilization and soap and water enough. what i mean by that is clean up my S.A.B. with soapy water leaving a nice puddle in the bottom of the S.A.B. then cleaning my agar jars with 70% iso putting them in my box then putting lid on wait for stuff to settle while shower come out ant go to work. clean jars again with soapy rag then flame sterilize my scalpel stick scalpel in agar then scrape spores onto agar. i did already sterilize the scalpel while i was pcing my agar. would in need to even flame sterilize since it is wrapped in foil to do 3 transfers of spores to agar?
i do know im not supposed to do iso and flame in a box just trying to wrap my head around the easiest method for sterilizing while working.
-------------------- want to grow like a Boss great links: How Frank gets shit done TL: HOW I DO EVERYTHING
Edited by j3rk0ff (10/01/13 08:21 AM)
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j3rk0ff
Stranger


Registered: 09/07/13
Posts: 80
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Re: agar help [Re: j3rk0ff]
#18915034 - 10/01/13 08:26 AM (10 years, 3 months ago) |
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-------------------- want to grow like a Boss great links: How Frank gets shit done TL: HOW I DO EVERYTHING
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hamloaf
Loaf of Fam.


Registered: 12/23/09
Posts: 20,192
Loc: Oklahoma.
Last seen: 1 year, 8 months
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Re: agar help [Re: j3rk0ff]
#18915037 - 10/01/13 08:27 AM (10 years, 3 months ago) |
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An inoculation loop is the preferred tool for handling spores. Scalpels are best used for cutting sections of already running mycelial culture that's on agar. Pcing the scalpel isn't necessary. Just wipe the scalpel with an alcohol soaked paper towel then flame sterilize the blade until red hot before each time the scalpel enters the SAB and makes contact the agar. Cool the scalpel's blade with unused agar medium before making contact with the culture. No need for a puddle of soapy water.
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retaardvark
Stranger

Registered: 12/22/12
Posts: 64
Last seen: 4 years, 8 months
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Re: agar help [Re: j3rk0ff]
#18915043 - 10/01/13 08:30 AM (10 years, 3 months ago) |
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You absolutely should not be wiping your jars with isopropyl; you're doing more harm than good. PC them if they are re-usable -- toss them and get new, sterile dishes if they are disposable. By opening and wiping them down, they are already hopelessly contaminated. Commercially-sterilized dishes are cleaned by gamma radiation, and iso ain't got SHIT on that.
Depending on how many dishes you're doing and the nature of the work, you should sterilize in between dishes or cuts. Some people, like myself, flame-sterilize between each and every dish. Others are comfortable enough to transfer a whole three-section petri dish to more without re-sterilizing. When in doubt, flame-sterilize more rather than less.
Never flame-sterilize in a closed box; it will ruin the still-air environment you've been cultivating. Remove utensils from SAB, wipe down gloves with iso paper towel, flame utensil, bring back inside while red hot.
edit -- WOOPS, thought you meant cleaning out the inside of your jars with iso.
Edited by retaardvark (10/01/13 08:31 AM)
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cynical bastad
another guy
Registered: 08/17/13
Posts: 175
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Post deleted by cynical bastadReason for deletion: .
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j3rk0ff
Stranger


Registered: 09/07/13
Posts: 80
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Quote:
I just used my right hand w/ tool being in SAB then stuck it out the Left hole and sterilized it then moved my other hand back in...it seemed to work better then bring both hand's out to flame and then both back in.
i like this idea thanks, im gonna have the wifey flame so i dont have to pull both out, but will keep this in mind when i dont have her around
Quote:
Also i read a thread the other day where RR was saying 17g agar to liter of water for germinating spores on a wetter medium of agar. So maybe cut your 10 back to 8.5ish?
i was concerned i didnt add enough agar. i used a syringe to dispense the agar to each agar jar. i then let them setup about 5 hrs before PCing. i touched one and it didnt seem like it was solid enough? 1st timer here so fingers cross.
-------------------- want to grow like a Boss great links: How Frank gets shit done TL: HOW I DO EVERYTHING
Edited by j3rk0ff (10/01/13 11:59 AM)
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FrankHorrigan
The Inquisition



Registered: 01/04/11
Posts: 10,573
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Re: agar help [Re: j3rk0ff]
#18916021 - 10/01/13 12:43 PM (10 years, 3 months ago) |
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I use 10g agar, 10g LME to 500mL of water. It works fantastic 
I'd recommend taking a look at my agar journal for a wide variety of recipes and agar discussion.
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cynical bastad
another guy
Registered: 08/17/13
Posts: 175
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Definitely check out Franks Journal Very cool stuff going on there.
But FH, if you were taking spores to agar, would you hydrate first?
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cynical bastad
another guy
Registered: 08/17/13
Posts: 175
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Re: agar help *DELETED* [Re: j3rk0ff]
#18916199 - 10/01/13 01:16 PM (10 years, 3 months ago) |
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Post deleted by cynical bastadReason for deletion: .
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j3rk0ff
Stranger


Registered: 09/07/13
Posts: 80
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just checked one after cooking they are good to go. im just concerned about the condensation on all that agar jars. gonna do some reading here in a few. hope it wont cause problems. gonna start working on the agar today.
-------------------- want to grow like a Boss great links: How Frank gets shit done TL: HOW I DO EVERYTHING
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cynical bastad
another guy
Registered: 08/17/13
Posts: 175
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Re: agar help [Re: j3rk0ff]
#18920148 - 10/02/13 09:38 AM (10 years, 3 months ago) |
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I don't know about the condensation, sorry
When your ready to try some Scorpions in your agar, hmu and i'll send some. Lol
Edited by cynical bastad (10/02/13 06:18 PM)
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j3rk0ff
Stranger


Registered: 09/07/13
Posts: 80
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will do thanks
-------------------- want to grow like a Boss great links: How Frank gets shit done TL: HOW I DO EVERYTHING
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