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By stry67
Posts:  99
Joined:  Mon May 02, 2011 6:54 pm
#112063
So I got this from Phyto labs because of the clarity recommendation. Product info says to use 2.5g per liter. I'm testing by adding "smidge" and "nip" amounts to 25ml in bfj's then autoclaving. Frankly I'm finding that it's gonna take a LOT more than 2.5g/L to get the right gel consistency. Anyone else use this and what have you found to be the right amount per liter?
By BradR
Posts:  450
Joined:  Sun Nov 28, 2010 5:00 pm
#112143
I use 2.75 g/l Gellan Gum for for CP media. Keep in mind that Gellan Gum needs divalent ions like ca++ or Mg++ to gel properly, unlike Agar. The more dilute the salts, the more Gellan Gum you need. 2.75 works fine for 1/3MS and 1/2MS media. If you were using full strength MS I'm sure 2.5g/l or even less would be fine.

Brad
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By Binny
Posts:  24
Joined:  Sun Jun 19, 2011 12:15 am
#112148
Take a look at Franks post here; http://www.flytrapcare.com/phpBB3/gelrite-t8300.html regarding pH and gelrite (sure different product slightly but same should hold true?).

Apparently there is a Gellan Gum mix with Agar that is the best of both worlds, but I haven't checked availability and have no experence except with agar myself. I am looking at getting something like this next order though.

Please update if you find a solution to this as I may be in the same boat shortly.
By stry67
Posts:  99
Joined:  Mon May 02, 2011 6:54 pm
#112315
Brad hit it on the head as for as what is needed to gel. I was trying to test it on just plain distilled water.Tho I find your 2.75g/L suspect. I did a batch today at 4g/L and it's just too soft. Yesterday i did a test batch at 8g/L and it was way too tough. Next round I'll run at 6g/L and pretty sure it's gonna be on the money or close to it. Gary from Phyto answered my email fairly quickly and stated using 1/3 MS, the 2g/L is gonna be nowhere close to waht's needed.
Having said all that, unless something un-foreseen comes up, I won't be going back to agar. Man, this stuff sets up like glass, I love it.
By BradR
Posts:  450
Joined:  Sun Nov 28, 2010 5:00 pm
#112496
stry67 wrote:Tho I find your 2.75g/L suspect. I did a batch today at 4g/L and it's just too soft.
stry67:
2.75 g/l is plenty of Gellan Gum. I am using the same G434 that you are using and I started out using 3g/l and it was a little too firm. I also use Phytagel, which is a Gellan Gum sold by Sigma and the results were the same. If you are requiring more than 3g/l with 1/2 MS media,then there is something else wrong with your technique. Possibilities are:
1. Your pH adjustment is wrong.
2. Your method of media preparation is wrong. I add the gelling agent last and always heat the media to the boiling point for a minute or two to make sure the gelling agent is completely dissolved. It will go from cloudy to clear when the gelling agent is completely in solution. Then the media can be dispensed into the vials and sterilized, or it can be sterilized in a media bottle and then dispensed into sterilized vials before it cools. Either way I swirl the vials to completely mix the gelling agent before it cools to the gelling point.

Brad
By stry67
Posts:  99
Joined:  Mon May 02, 2011 6:54 pm
#112549
BradR wrote:
stry67 wrote:Tho I find your 2.75g/L suspect. I did a batch today at 4g/L and it's just too soft.
stry67:
2.75 g/l is plenty of Gellan Gum. I am using the same G434 that you are using and I started out using 3g/l and it was a little too firm. I also use Phytagel, which is a Gellan Gum sold by Sigma and the results were the same. If you are requiring more than 3g/l with 1/2 MS media,then there is something else wrong with your technique. Possibilities are:
1. Your pH adjustment is wrong.
2. Your method of media preparation is wrong. I add the gelling agent last and always heat the media to the boiling point for a minute or two to make sure the gelling agent is completely dissolved. It will go from cloudy to clear when the gelling agent is completely in solution. Then the media can be dispensed into the vials and sterilized, or it can be sterilized in a media bottle and then dispensed into sterilized vials before it cools. Either way I swirl the vials to completely mix the gelling agent before it cools to the gelling point.

Brad
I am using 1.46 g/L of MS which works out to roughly 1/3 MS. I said I thought 4g/L was too soft but actually it seems about right. That amount is inline with what Frank Tromble and Gary from Phytotech told me to use based on greatly reduced levels of catians. My ph coulda been off a bit since I was using 2 month old buffer solution to calibrate my meter but I have new solution coming. So the next batch of MS I will be able to do a more accurate ph reading I'm sure. Speaking of ph, I asked Gary about it and he said they (Phytotech) have not seen any interaction between G434 and ph. That G434 seems to work regardless of ph, that they have customers who use G434 in very acidic media where even agar wont gel.
My media preparation is inline with yours except I didn't find the need to have it actually boiling. The second round I slowwwwwwly added the gum while vigorously stirring and so there was no clots.
By BradR
Posts:  450
Joined:  Sun Nov 28, 2010 5:00 pm
#112618
Here is a tube of media gelled with 2.75 g/l Gellan Gum G434, Lot number11A043-4024A. The media is 1/2 MS with a pH of 5.7. The consistency is perfect. No sagging when tilted or tapped, but soft enough for the roots (Wacky Traps) to easily penetrate. One of the coolest thing about using clear media is being able to see those beautiful roots.

Brad
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By TCGator
Posts:  44
Joined:  Fri Jan 16, 2015 9:21 pm
#322630
Even with the cold-pour method that many labs use, 4 grams per liter of gellan gum would be overkill to solidify the medium that Phytotech offers as a "pretransplant" medium.
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