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Offlinear1es
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Registered: 07/09/10
Posts: 776
Last seen: 2 years, 2 months
Crapppp.... Its whiskey time!
    #14089989 - 03/08/11 11:22 PM (13 years, 10 months ago)

So i finally got around to doing some real agar work.

I used agar before but just threw a bit in the bottom of 1/4 pint jars

it wasnt half bad actually because id pour the jars with agar then sterilize

once they were done i kept in fridge until i was ready to use

whenever i saw a cluster-y pin id hack it off and throw it in a jar

pretty cool stuff

its hard to see in the jars on what is going on but you can at least see growth through the bottom

ive seen others that used saran wrap for lids

my lids just had a bit of polyfill or tyvek as filters

i had some decent luch with the jars, would open them up, cut a small piece then transfer to other jars

then when these grew out a bit id slice up and throw in jars of grain

i admit i was a little lazy and left some jars go a bit too long and lo and behold they pinned fruited and dropped spores lol


but i digress

i wanted to get to doing isolations so i picked up some of those 3 section Y-petri dishes and some PDA

mixed up 24 grams of PDA in 600 ml water using a quart jar as the container (first mistake) placed a tinfoil lid on top of my tyvek filtered lid

pressure cooked for about 40 minutes

my AA tends to get a bit hot on my stove and it always creeps past 15 psi up over 20 psi so im pretty sure i cooked the hell out of the pda

waited forever for the pda to cool down to usable temps

so i prepared my glovebox.

i have a small hepa filter with a UV-C light that i was running in my glovebox room

i put all my supplies in the glovebox and was waiting for my pda to cool enough to pour


when it was ready i began my quest

first off i thought my glovebox was pretty damn good

its got 2 holes for the arms with those couplings built into em

i used to have gloves attached to the glovebox but i got rid of those because they restricted my movement too much and now just use tyvek sleeves and latex gloves

i got a plexyglass lid thing going on so i thought id be golden but it wasnt big enough

add to that the fact that the bottom of the glovebox isnt completely flat

its kinda convex

i opened up the sleeve of petri dishes and i set them in the corner of the glovebox

then i opened up the lid of the quart jar to prepare to pour my jars

when i sat down the quart jar the weight of the jar pushed down on the convex bottom of the box and it popped my petri dishes up like being double bounced on a trampoline lol

so i go to start pouring the agar and i cant see shit

well thats not entirely true

i can see from top down but that doesnt help me gauge how much agar im pouring into each section of petri, sometimes the section would get too little, sometimes too much

also looking from top down i could see the agar from the dishes id already poured through the bottom of each new dish. also made it difficult to tell how much was being poured


for my first petri pour i should have probably just went with regular petri dishes

using the Y section petri dishes meant I had to pour 3 different times for each dish.

it was so fucking frustrating

then i noticed the clumps

apparently i waited a bit too long and when i opened the jar it accelerated the cooling process so the 2nd set of 10 petri dishes were a goddamn mess. id get a little agar into the section, then a big chunk would land, spilling agar over the sides

i ended up having 4 unpoured petri dishes left over when i was done so i put them back in the sleeve and hopefully they will be good for the next round of agar

anyway what the the hell

like most things in this hobby

the first time i do it im a complete idiot but then i get it down until i can do it hungover, half asleep and stoned all  while talking on the phone no problem

im going to take RRs advice on using a liquor bottle because it seems to make the pour a lot easier

I recently read someone using one of those

jesus-watching "I Saw The Devil" atm... crazy shit

i read someone using those liquor pour things that you see bartenders use to help pouring agar as well

im also going to have to either add a side window to my glovebox or just nix the lid i fashioned and replace it with a huge piece of plexyglass as per RoadKill's glovebox

ah well, im waiting for the plates to cool now while watching this movie and will be inoculating them with some cube spores

im going to try and wipe up any mess i may have made on the sides, wrap them bitches in parafilm and then hope for the best

as they say, every cloud has a silver lining

i get to go to the store tomorrow and pick up a bottle of whiskey and down it with a few friends to make my next go round a bit easier

gotta love mycology!


--------------------
":mushroom2:here is nothing lost or wasted in this life.” - Bhagavad Gita

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Offlinear1es
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Registered: 07/09/10
Posts: 776
Last seen: 2 years, 2 months
Re: Crapppp.... Its whiskey time! [Re: ar1es]
    #14090533 - 03/09/11 01:07 AM (13 years, 10 months ago)

updated

it didnt turn out too bad

just finished inoculating with a cube print and wrapping with parafilm

i wasnt sure if i should keep all the parafilm inside of the GB or not

i ended up keeping it ouside of the box and grabbing a piece between each dish

i bought 2 inch parafilm which means that it is about 2 "squares" across when i pull it out of the box

i read that i should use 4 squares of parafilm for each plate so id pull 2x4 squares out at a time and then cut it in half lengthwise to get the 1x4 square pieces that i need.

i did do this before hand

when using parafilm its not like regular tape

i should have figured it because the came is parafilm so its this waxy stretchy tape

it seems as if there is a sweetspot in terms of how much it needs to be stretched before it makes sweet sweet love to the petri dish and hangs on for dear life

good god this movie is graphic

i think the 1x4 row of parafilm may be a bit much but its cheap so what the hell. RR hasnt led me wrong so far lol

looking back i probably should have kept the tape inside

ah well

i think the room was pretty clean though
i had a hepa filter running and kept spraying oust periodically

plus i had gloves on and tyvek sleeves

so

after inoculating and wrapping these badboys i finally got to take a look

of the 16 y shaped petris i poured, probably 6 of them were a either a bit heavy or a bit light on the agar

a couple of them had so much agar that the lids stuck and i had to pry them apart crappppp

i had enough sections poured properly that i should be aight

hopefully my snafu with the parafilm wasnt enough to contam my experiment

next time ill make sure i can see what im doing, have a bottle thats easier to pour, and keep the parafilm inside the still air box so i dont have to remove my arms once they are inside

i apologize for not having any pics on this post. ill go borrow a digital camera and try and update it so it doesnt suck as much

also plan on posting the results of my agar experience if and when i see growth


--------------------
":mushroom2:here is nothing lost or wasted in this life.” - Bhagavad Gita

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Offlinebbt0009
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Registered: 11/11/08
Posts: 542
Last seen: 12 years, 19 days
Re: Crapppp.... Its whiskey time! [Re: ar1es]
    #14090569 - 03/09/11 01:19 AM (13 years, 10 months ago)

too drunk too read :ilold:

Its bookmarked for tomorrow bro im sry


--------------------
It was all a dream...

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OfflineHitsuzen
Space Cowboy
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Folding@home Statistics
Registered: 10/17/08
Posts: 399
Loc: Earth Flag
Last seen: 4 years, 1 month
Re: Crapppp.... Its whiskey time! [Re: bbt0009]
    #14090603 - 03/09/11 01:30 AM (13 years, 10 months ago)

damn dude I'm drinking whiskey and I totally just drunk-Facebook'd 5 ex-girlfriends. the weed probably didn't help either. cheers! :awehigh:


--------------------
Hate never yet dispelled hate,
Only love dispels hate.
This is the law,
Ancient and inexhaustible.
-Buddha


                           

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