Tips For Transfering Agar Wedges?


Nan's Nook : Archives : Cloning : Agar
Posted by: DirtyWOP Jan 23 03, 12:31 PM GMT
Sticky agar.....thats right.
Everytime I try to transfer a wedge I have a problem with it sticking to everything......either the scalpel or the side of a jar...and I ruin cultures by wasting time, leaving them open and klutzing around trying to get the wedges where I want them....

I remember reading somewhere that there is something wrong if your agar sticks to everything....is that correct?

I think I just need to use a long-ass needle to move it around....
if anyone has any tips, thanks
if not, no biggy

Posted by: DirtyWOP Jan 23 03, 01:06 PM GMT
well...
Paul stamets says that is because my agar is too thin....
which isn't the case....

Also, anyone know if you can caramalize malt agar, and is that bad? I have heard it IS bad, and that it IS good.......never know who to believe anymore.......

Posted by: Zoom Jan 23 03, 01:19 PM GMT
Hey Wop...My agar does the same thing and I make it as directed. Malt agar will carmelize. Not as effective when it does because sugars are not evenly distributed. Has a crystalizing affect. That's all I know buddy but I'm sure you'll get more posts.

Posted by: Mycota Jan 23 03, 01:34 PM GMT
You might give PDA agar mix, a try.

I found some "Exacto" knife type scaplel looking things at Home Depot (by the cash registers, on racks). They are sort of pen sized, sort of for wood carving, come with replacement blades & work perfectly for cutting & transferring agar wedges.

Mycota.

Posted by: DirtyWOP Jan 23 03, 01:46 PM GMT
I don't think it will carmalize if you PC at 10 lbs of pressure.......just was wondering if carmalized sugars will inhibit healthy growth....

Yea mycota, I know....I just have way too much pre-mixed malt agar and I wanna use it, it was like 25 buck (prolly a waste of money) . I also have some dextrose and agar agar, so I'm GONNA do the potato thing, just not yet.....not to mention all the potato broth liquid cultures I've tried got NASTY....as opposed to ones with malt which are still good....I think malt is less prone to contam.

Yea, I've got a few x-actos scalpels but the agar is still sticky and still sticks to the blade...

Wop's shoes:
imagine you got your wedge sliced, you pick it up with you scalpel, open the sterile jar or pitri, and when you try to drop the wedge in there, it just don't drop. Then you start banging it on the rim of the jar of pitri and it still don't drop. Then you end up touching the agar surface with your tool(which I don't like to do) trying to get it off and it still don't drop. And all the while spores are flying all over the place, and your getting all frustrated........then you get it to finally fall in, and it sticks to the glass....
you getting the mental picture?
it sucks

I'm thinking a thin needle will solve my problem. If I only pick up the wedge with the very tip of a needle, it is barely hangin there, and should drop easily.....I guess
I'll try it next time.....

Posted by: DirtyWOP Jan 23 03, 01:48 PM GMT
Also....if anyone knows any good agar mixes for woodlovers like reishi I would love to hear em.....can I make a "tea" out of woodchips and cardboard then add dextrose?

...and at what temp should reishi be incubated? In the fridge?


*Man, this guy needs to shut up and do some research, huh?....


Posted by: Mycolaureat Jan 23 03, 03:34 PM GMT
Malt agar is too sticky and unclear ...

Potato and dextrose agar is good for all lot of species including some woodloving species like ps.cyanescens

Posted by: DirtyWOP Jan 23 03, 03:46 PM GMT
thanks man
it IS a bitch to spot bacterial contams with that murky color and all the undissolved solids.......

o well, I'll use it up eventually.....
better to have it than not

Posted by: G a n j a Jan 23 03, 06:17 PM GMT
Dark malt syrup has all ready caramelized a lot of the sugar content ware as light malt that comes in powder form has more available sugars.

Also try using 2 tools,one in each hand for transpheres so you can
nudge one of with the other.

Good luck

P e a c

Posted by: DirtyWOP Jan 23 03, 06:31 PM GMT
nah
I can't hold two tools
I only have one available hand
because I'm usually holding the lid of the jar or pitri in the other
so I can quickly put it back on....

Posted by: OneDiaDem Jan 23 03, 06:52 PM GMT
QUOTE
Wop's shoes:
imagine you got your wedge sliced, you pick it up with you scalpel, open the sterile jar or pitri, and when you try to drop the wedge in there, it just don't drop. Then you start banging it on the rim of the jar of pitri and it still don't drop. Then you end up touching the agar surface with your tool(which I don't like to do) trying to get it off and it still don't drop. And all the while spores are flying all over the place, and your getting all frustrated........then you get it to finally fall in, and it sticks to the glass....
you getting the mental picture?
it sucks


That was not really something to laugh at Wop, however, it did hit my funny bone hard, only cause I have been there many times.

If you are working in a glovebox, and have pc'd everything, here is something that may help.

When you load your pc, fold up a peice of foil about 12"x12" all by itself. Sterilize 2 blades, or a blade and a small screwdriver or something in another peice of foil.
When you get in the glovebox, use the 12x12 foil to lay your lids on so you have both hands free.
I use a screwdriver that looks like a ice pic to push the agar into the grains off the scalpel.

When I have tried to hold the lid, it just goes downhill from there. So, I am with ya on the frustration levels.



Posted by: Fungusmaximus Jan 23 03, 07:05 PM GMT
uuhhh... Tweezers anyone? if it sticks, then give it a little jolt and it will go.
I just stab it with the exacto and use the agar to pull it off. My first transfer ever I was a bit stoned and using open arm holes, for some reason I stuck my finger in the agar trying to get a wedge out(not thinking)
I realized what I did and a fury of cussing followed. I thought for sure the culture was contam'd, nope nothing but myc biggrin.gif I have the same culture in the fridge to this day, still going strong smile.gif

Posted by: OneDiaDem Jan 23 03, 07:34 PM GMT
Tweezer are ok. I used them once. I like the ease of being able to wipe a poker off with a cotton ball soaked with alcohol tho. I have a hard time cleaning the tweezers with gloves on.

Posted by: DirtyWOP Jan 23 03, 08:32 PM GMT
Yea, I used tweezers today to transfer mushroom tissue to a jar - went pretty smooth.....it's good for getting nice tiny specks (less contams that way), but I must say, the chunk DID stick to the tweezers.....and I had to tap it a few times on the edge of the jar, and the chunk didn't land exactly in the center....but it'll be ok.


Posted by: Fungusmaximus Jan 23 03, 10:23 PM GMT
Shit sometimes I roll those chunks around inside the culture dish/ jar if Im just trying to colonize the plate.

Posted by: tron Jan 23 03, 10:26 PM GMT
hell wop, i bet if you ate something the shakes would go away and your transfers would be a smooth as nans ass.

Posted by: Molester Jan 23 03, 10:34 PM GMT
The icepick screwdriver is a good idea. The less surface area on the secondary tool, the less grip for the agar.

I just use a pinch of agar with a couple pinches of dextrose in a 100 mL of water... works beautifully, seems to give the dextrose tek a good bump. Growth everywhere in no time. Haven't tried it on woodlovers yet though. But its so easy to make, and so easy to suck it up and shoot it.

Posted by: DirtyWOP Jan 23 03, 11:09 PM GMT
hehe tron, you're funny!

Indeed molester!
I just tried out a long pushpin playing with the TX strain, and it worked great....illustrates your premise there. But despite the fact that the wedges are easy to DROP with the pin, conversely its harder to pick them up...tongue.gif......I guess you win some, you lose some......


Not sure I get the picture tho dood....
Why shoot agar? It's only a solidifying agent.....
seems like you'd be better off without it.... huh.gif

Posted by: Molester Jan 24 03, 12:14 AM GMT
Shit, are you serious? I thought agar had nutes in it. Its made from seaweed. Maybe I'm gettin better growth, simply because I'm using less dextrose? It SEEMED to be helpin! I'm gonna hafta take another look at the jars now tongue.gif Yeah, lookin more beautiful than ever, by far!

Posted by: DirtyWOP Jan 24 03, 12:34 AM GMT
No nutes in the agar itself, thats why you have to add malt, or potato broth and dextrose.....

I don't see how it was so easy to suck up and shoot....the purpose of agar is to solidify! Maybe only adding a pinch kept it fluid.....?

Posted by: Molester Jan 24 03, 12:55 AM GMT
Hmnnn. Some of the agar solidified at the bottom, but it also made a nice suspension. So maybe the benefit was suspending the spores? Worked wonders! My dextrose jars always sucked ass till I started this. This way the spores really reach for their brothers and sisters, giving a lot more favor to the rhizos, because they get to fuck way first. Cottony growth is yet to be seen in my jar. Tried to get a picture, but the glare on the glass, the texture of the jar, and the tint of the agar screwed it up. And I dropped the jar on its side while putting it back! I hope the liquid didn't get contamed from the filter patch. cry.gif cry.gif cry.gif

Posted by: Fungusmaximus Jan 27 03, 06:45 PM GMT
What about thinly coating your forceps w/ some parrafin wax?
Heat up the wax dip em quickly . Dry time is only a few seconds, and its clean. Shit wont stick to the wax.

Posted by: prisoner#1 Jan 28 03, 01:17 PM GMT
Another suitable method is a mineral oil dip. Nex time you autoclave anything include a pint or two of mineral oil. You can use it fo a multitude of purposes. topping colonized agar slants to slow the metabolism and coating instruments to prevent things sticking, like agar

Prisoner#1

Posted by: DirtyWOP Jan 28 03, 01:21 PM GMT
great responses, thanks....

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