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No Tilt Agar Jars......


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#1 Microbe

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Posted 15 June 2014 - 05:18 PM

I decided to use my cheap version of the No Tilt LC jar to improve the process of pouring plates and or slants and I must say it work beautifully. IMO this has to be the cleanest and most efficient way of pouring plates or slants of all the methods I have tried.

What I did.....

Prepared the volume of nutrient agar that I needed using the method I have always used.

Dump it into my No Tilt Jar and PC for 25 minutes.

asaqagyt.jpg

I prepped and cleaned my working area and laid out all the instruments that I will be using and these also were cleaned.

ynezy5um.jpg

Used an airport syringe to remove the vacuum created after the PC run.

4ugu7aje.jpg

Next I laid a paper towel soaked in ISO over the lid and unscrewed male luer lock cap from the airport on my NTJ.

ne8yvyhe.jpg

Then I pulled the paper towel back enough to attach the airport syringe like so.

7e5y7ega.jpg

Did the same for the withdraw port. I removed the paper towel for capturing an image. I leave it there so when I remove a full syringe I can cover the port.

aty6e4uv.jpg

I draw the nutrient agar.

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I put the paper towel back so that when I detach the syringe there is a barrier to reduce the risk of contamination.

enynysan.jpg

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Attach a luer lock syringe cap and allow to cool to the target temperature and consistency for pouring or in this casing injecting.

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Repeat the process to fill as many syringes that you need and or until the nutrient agar is depleted.

I filled 10 plates in approximately 1 minute!

asu9u6y5.jpg

So after I did this I thought about making many syringes to have agar on demand. Couldn't I make up several pints of of agar and draw it into a syringe using this method and store it until needed? I could pop it in the microwave to liquify it when I need it. Maybe even freeze it assuming I left some air in there for it to expand. What do you guys/gals think about preparing a lot of syringes filled with nutrient agar for agar on demand?
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#2 Microbe

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Posted 16 June 2014 - 05:47 PM

Has anyone ever froze their agar? I was thinking about making up about 25 30 cc syringes full of PDA to have on demand so that I can pour a few plates at a moments notice and not have to pour so many at once if I don't need them. I typically pour 20-40 plates at a time with the minimum being 10, but I don't always need so many all at once spending on what I'm working on but would love to pour a single plate if I so chose to do and thought that storing them in the freezer would work or maybe even the fridge......

#3 wharfrat

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Posted 16 June 2014 - 08:21 PM

i have never tried freezing agar, i have tried to freeze jello, which is similar, and it turns to a gunky water when thawed. Nice 'no tilt' setup. do you like the black agar? i have never seen black. vs. a clear'sh agar that u can see both sides of the dish.



#4 the_chosen_one

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Posted 16 June 2014 - 08:34 PM

i like the black/dark for isolating monokaryon mycelium. much easier to see using a magnifier ime.

adding food coloring to the agar works too. flavored syrups optional :wink:

 

agar can be stored in the fridge and re-liquified. i have only heard bad about freezing it. as wharfrat noted with the jello. the freezing extracts the water from the agar and you have a mess. doesn't merge well after. 

 

and also, very nice write up and beautiful lab! the black latex with the black agar against the white background is a real turn on. 


Edited by the_chosen_one, 16 June 2014 - 08:41 PM.

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#5 wharfrat

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Posted 16 June 2014 - 08:39 PM

i like the black/dark for isolating monokaryon mycelium. much easier to see using a magnifier ime.

adding food coloring to the agar works too. flavored syrups optional :wink:

cool. my agar work has been very limited. as i am using a positive pressure glovebox vs. a flowhood. very cumbersome. and my dishes are the 1/2 pint flat jelly jars, w/out clear lids, so i kinda need a clear agar. black would look pretty cool with a real petri dish


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#6 the_chosen_one

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Posted 16 June 2014 - 08:45 PM

 

i like the black/dark for isolating monokaryon mycelium. much easier to see using a magnifier ime.

adding food coloring to the agar works too. flavored syrups optional :wink:

cool. my agar work has been very limited. as i am using a positive pressure glovebox vs. a flowhood. very cumbersome. and my dishes are the 1/2 pint flat jelly jars, w/out clear lids, so i kinda need a clear agar. black would look pretty cool with a real petri dish

 

i use the clear for most things. especially no pour agar teks. the dark is just nice for breeding purposes.



#7 Microbe

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Posted 16 June 2014 - 08:56 PM

i have never tried freezing agar, i have tried to freeze jello, which is similar, and it turns to a gunky water when thawed. Nice 'no tilt' setup. do you like the black agar? i have never seen black. vs. a clear'sh agar that u can see both sides of the dish.


I add food coloring to turn it black. I tried it out and it really makes seeing any germination very easy. Molds, bacteria, and if course mycelium. Not that I was struggling seeing it with the regular agar , just stating I see it sooner on the black.

Thanks for the kudos by the way.
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#8 Microbe

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Posted 16 June 2014 - 09:00 PM

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

#9 wharfrat

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Posted 16 June 2014 - 09:01 PM

 

i have never tried freezing agar, i have tried to freeze jello, which is similar, and it turns to a gunky water when thawed. Nice 'no tilt' setup. do you like the black agar? i have never seen black. vs. a clear'sh agar that u can see both sides of the dish.


I add food coloring to turn it black. I tried it out and it really makes seeing any germination very easy. Molds, bacteria, and if course mycelium. Not that I was struggling seeing it with the regular agar , just stating I see it sooner on the black.

Thanks for the kudos by the way.

 

i wanna try it!!  i gotta get some real dishes. <--- noob ;)


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#10 wharfrat

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Posted 16 June 2014 - 09:04 PM

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

i leave mine in the fridge, but u can only heat it back up so many times.. 3 times for bulk and i throw it away, it starts getting darker in color. not sure if that affects the nutrition but as i stated ^ above, i need to be able to see through it.



#11 the_chosen_one

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Posted 16 June 2014 - 09:06 PM

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

your only issue might be remelt. i've never seen it done with syringes. only jars. but if the syringes are packaged in the clean area i don't see why not. leave some room for expansion and heat using a double boiler so you can see. might have a blow out or two getting dialed but my hunch is it will work. 


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#12 the_chosen_one

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Posted 16 June 2014 - 09:08 PM

 

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

i leave mine in the fridge, but u can only heat it back up so many times.. 3 times for bulk and i throw it away, it starts getting darker in color. not sure if that affects the nutrition but as i stated ^ above, i need to be able to see through it.

 

yeah it will eventually mess up sugars. i find vanilla extract to be very resilient to heat. 


Edited by the_chosen_one, 16 June 2014 - 09:09 PM.


#13 Microbe

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Posted 16 June 2014 - 09:08 PM

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

i leave mine in the fridge, but u can only heat it back up so many times.. 3 times for bulk and i throw it away, it starts getting darker in color. not sure if that affects the nutrition but as i stated ^ above, i need to be able to see through it.

Yeah if I heated my syringe up I would dispense the entire contents probably. 35 cc syringe will fill 3 dishes to the volume I like.

The link below is the cheapest place I have found plates that don't requiring you to wait 3 months for them to arrive. I have 160 of the 60 mm and I love them. Hold them in 1 hand and manipulate the cover with the thumb.

http://www.scientifi...tri-dishes.html

#14 wharfrat

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Posted 16 June 2014 - 09:11 PM

bookmarked, thanks!



#15 wharfrat

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Posted 16 June 2014 - 09:11 PM

cheaper then what i am using



#16 Microbe

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Posted 16 June 2014 - 09:18 PM

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

your only issue might be remelt. i've never seen it done with syringes. only jars. but if the syringes are packaged in the clean area i don't see why not. leave some room for expansion and heat using a double boiler so you can see. might have a blow out or two getting dialed but my hunch is it will work.

Would 5cc be enough room you think for expansion?

#17 wharfrat

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Posted 16 June 2014 - 09:25 PM

 

 

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

your only issue might be remelt. i've never seen it done with syringes. only jars. but if the syringes are packaged in the clean area i don't see why not. leave some room for expansion and heat using a double boiler so you can see. might have a blow out or two getting dialed but my hunch is it will work.

Would 5cc be enough room you think for expansion?

 

you would have to have them harden and melt standing on the plunger, so the air is at the needle point, or it will blow out when it melts



#18 Microbe

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Posted 16 June 2014 - 09:27 PM

cheaper then what i am using


I have not shopped their polyethylene Petri dishes but I'm sure they are economical as well. I don't like them because they will melt in a PC leaving them disposable and 1 time use only unless you want to attempt sterilization with chemicals. I don't know about you but I have had 60 plates going at one time and have read that isolating could take 100 + plates. But man it sucks big time washing them. I just washed 40 the other day that had been hanging around in a bucket!

HATED IT!!!

9e8ymute.jpg

I'm no longer transferring multiple selections of mycelium. I'm doing it 1 plate at a time so that I do to double my cultures unless attentions are to increase inoculant. I would start with 4 plates then 8, 16, and etc.. Became overwhelming but was able to be very selective on the myc that I put into my jars on my B+ grow. I love agar work just hating cleaning up after it and disposable plates would be to expensive for my needs.
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#19 the_chosen_one

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Posted 16 June 2014 - 09:29 PM

 

 

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

your only issue might be remelt. i've never seen it done with syringes. only jars. but if the syringes are packaged in the clean area i don't see why not. leave some room for expansion and heat using a double boiler so you can see. might have a blow out or two getting dialed but my hunch is it will work.

Would 5cc be enough room you think for expansion?

 

maybe start at half of a syringe, maybe two thirds and work up. i doubt you'll need to actually boil the agar. i usually melt a jar in the pc on low for 10 minutes and i think that is overkill. the syringes will have to be handled gently. could be a bit dangerous. be careful. did you have caps for them?.. haven't looked yet.. i won't even say why this time lol. anyways i would probably tape the plunger in place. that way the tip can be pointed away. 



#20 Microbe

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Posted 16 June 2014 - 09:29 PM

So it is safe to say that I could make up a bunch of agar syringes and store in the fridge to have agar on demand? That just really made my day because there were so many times I wanted to transfer i piece of mycelium that came out of nowhere or attempt to clone but didn't have any agar plates poured. I almost always have 20+ plates going but that's just it, they always have something in them.

your only issue might be remelt. i've never seen it done with syringes. only jars. but if the syringes are packaged in the clean area i don't see why not. leave some room for expansion and heat using a double boiler so you can see. might have a blow out or two getting dialed but my hunch is it will work.
Would 5cc be enough room you think for expansion?
you would have to have them harden and melt standing on the plunger, so the air is at the needle point, or it will blow out when it melts

Thank you. I'm going to make some up in the next few days .




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