|
 joehinkle
JustLearning
Registered: 11/02/08
Posts: 132
Loc: USA
Last seen: 3 years, 7 months
|
 Shake agar prior to Solidifying 
#10200265 - 04/20/09 10:23 AM (4 years, 1 month ago) |
|
|
I bought some premixed agar from FP.
When test tubes (agar + wood) for slants comes out of the PC, the mixture is separated with some sediment on the bottom.
The first set I made, I shook prior to letting them solidify but ended with agar bubbles populating the top to the slant.
The second set I made I did not shake and left the sediment at the bottom.
When using agar, should they be shaken just before allowing them to solidify and live with the bubbles?
Edited by joehinkle (04/20/09 10:33 AM)
|
solumvita
Q.B.E.
Registered: 02/12/08
Posts: 1,975
Loc: South Africa
Last seen: 18 days, 10 hours
|
Re: Shake agar prior to Solidifying [Re: joehinkle]
#10201025 - 04/20/09 01:04 PM (4 years, 1 month ago) |
|
|
Are you dissolving the agar in boiling water before you put it into the PC?
--------------------
One of these days all the answers will be revealed until then we learn from each other!
www.mushrush.co.za
|
 lipa
Registered: 07/24/07
Posts: 1,876
Loc: USA 
|
Re: Shake agar prior to Solidifying [Re: joehinkle]
#10201102 - 04/20/09 01:20 PM (4 years, 1 month ago) |
|
|
I just twirl them a little when I set them down to solidify. They are usually pretty darn hot still when I do that. A little sediment is ok. I cant believe you are shaking culture tubes with agar in them. By bubbles you mean a bunch of agar all over the side of the tube. Like you can't see inside them.
Lipa
--------------------
John Landis Mason is my hero 
|
joehinkle
JustLearning
Registered: 11/02/08
Posts: 132
Loc: USA
Last seen: 3 years, 7 months
|
Re: Shake agar prior to Solidifying [Re: lipa]
#10201383 - 04/20/09 02:34 PM (4 years, 1 month ago) |
|
|
The directions on FP's agar does not state that the agar needs to be boiled prior to placing into a permanent container - it only states to PC it.
RR's video does not boil prior to pouring into test tubes.
I searched the forums and got a mixture of suggestions to boil and not to boil.
I was concerned about the sediment with the first set of bubes so I shook it after removing it from the PC. Still hot - but it formed bubbles.
Here is a pic - I have already seeded it with a slice of agar mycelium.
I have 7 more cultures to add to slants so last night I made up 40 new tubes.
I did not shake this time - since RR did not shake them on his video.
The pic below is the worst of the lot as far as sediment. My only concern is the composition of the agar mix for extending the life of any culture I place it them.
Before I seed these 40 slants with mycelium (5 slants per mushroom culture) I just want to make sure I'm not starting out with garbage.
Thanks in advance for your comments.
|
Jef
Out-of-work Sex Slave
Registered: 12/03/08
Posts: 739
Loc: near Duncan, BC
|
Re: Shake agar prior to Solidifying [Re: joehinkle]
#10203836 - 04/20/09 10:35 PM (4 years, 1 month ago) |
|
|
I always boiled. I did MEA.
I always had sediment too, and consider that normal. Of course the last plates had more sediment than the first.
There was no performance difference that I could discern.
I think you're all okay.
--------------------
I am my own lab rat.
“Young lady!” cried Mr. Grout, very much appalled. “It ill becomes a young person of no consequence, such as yourself, to go about slandering great people in all the dignity of their property!”
- Mrs. Mabb, The Ladies of Grace Adieu, by Susanna Clarke
|
joehinkle
JustLearning
Registered: 11/02/08
Posts: 132
Loc: USA
Last seen: 3 years, 7 months
|
Re: Shake agar prior to Solidifying [Re: Jef]
#10208884 - 04/21/09 07:45 PM (4 years, 1 month ago) |
|
|
Lesson Learned.
The unshaken slants looked OK ... so I setup the flow hood and started to do some transfers from dish to slant.
As I placed a sample in the slant I noticed a small amount of liquid.
I looked at all 40 slants and 80% have liquid which means they did not cure correctly ....
Now I'm disposing of all 40 slants and starting over.
I did not know you could get a partial solidification with agar ...
That sediment on the bottom - (I shook and mixed with the first set of slants and they came out ok) must be part of the solidification reaction.
Hard lesson to learn (time spent wise)!
|
RogerRabbit
Bans for Pleasure
Registered: 03/26/03
Posts: 39,269
Loc: USA Mountain Northwest
Last seen: 8 days, 2 hours
|
Re: Shake agar prior to Solidifying [Re: joehinkle]
#10210253 - 04/21/09 11:02 PM (4 years, 1 month ago) |
|
|
I always shake the bottle like hell prior to sterilization and before filling the slants. That tends to reduce the sediment. I also make up the FP agar about 20% weaker than called for in the instructions. This takes care of most of the sediment and the slants solidify normally.
When pouring plates, I always give the bottle of agar a good swirl just before pouring. It also helps to let the agar cool nearly to the point it's starting to solidify before swirling and pouring.
RR
--------------------
Download Let's Grow Mushrooms
semper in excretia sumus solim profundum variat
|
solumvita
Q.B.E.
Registered: 02/12/08
Posts: 1,975
Loc: South Africa
Last seen: 18 days, 10 hours
|
Re: Shake agar prior to Solidifying [Re: joehinkle]
#10211206 - 04/22/09 01:44 AM (4 years, 1 month ago) |
|
|
Quote:
joehinkle said:
As I placed a sample in the slant I noticed a small amount of liquid.
I looked at all 40 slants and 80% have liquid which means they did not cure correctly ....
most likely you poured the agar when it was too hot, that causes condensation and accounts for the water in the tubes, only time i get partial solidification is when i make "soft water" agar and that is on purpose.
--------------------
One of these days all the answers will be revealed until then we learn from each other!
www.mushrush.co.za
|
Jef
Out-of-work Sex Slave
Registered: 12/03/08
Posts: 739
Loc: near Duncan, BC
|
Re: Shake agar prior to Solidifying [Re: solumvita]
#10211241 - 04/22/09 01:51 AM (4 years, 1 month ago) |
|
|
If solumvita is correct, you can just pour off the excess.
--------------------
I am my own lab rat.
“Young lady!” cried Mr. Grout, very much appalled. “It ill becomes a young person of no consequence, such as yourself, to go about slandering great people in all the dignity of their property!”
- Mrs. Mabb, The Ladies of Grace Adieu, by Susanna Clarke
|
solumvita
Q.B.E.
Registered: 02/12/08
Posts: 1,975
Loc: South Africa
Last seen: 18 days, 10 hours
|
Re: Shake agar prior to Solidifying [Re: Jef]
#10211961 - 04/22/09 05:19 AM (4 years, 1 month ago) |
|
|
that's exactly what I do
--------------------
One of these days all the answers will be revealed until then we learn from each other!
www.mushrush.co.za
|
joehinkle
JustLearning
Registered: 11/02/08
Posts: 132
Loc: USA
Last seen: 3 years, 7 months
|
Re: Shake agar prior to Solidifying [Re: solumvita]
#10214740 - 04/22/09 07:00 PM (4 years, 1 month ago) |
|
|
As it turned out - there was NOTHING wrong with the slants.
It was condensation I was seeing.
Live and learn by our mistakes!
|
|
Images only 
Threaded
Previous
Index
Next
Shop for: 
Edit 
Reply 
