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mushhiehunter
Trichodermatologist


Registered: 10/11/16
Posts: 837
Loc: Česká republika
Last seen: 3 years, 2 months
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TIFU agar soaked in LC
#23739209 - 10/15/16 05:42 AM (7 years, 3 months ago) |
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Hey guys, it was my first time doing LC to agar and I messed up, I couldn't see properly through the glovebox and I squirted quite a bit of LC in the jars, now should I let it be and avoid potential contamination or should I take a sterile needle and suck back up the excess liquid?
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tump
ban the undead



Registered: 03/17/16
Posts: 2,383
Last seen: 6 years, 10 months
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Start over again. And let it be if you have the spare jars. You are injecting lc to agar to test the clean of the lc
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mushhiehunter
Trichodermatologist


Registered: 10/11/16
Posts: 837
Loc: Česká republika
Last seen: 3 years, 2 months
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Re: TIFU agar soaked in LC [Re: tump]
#23748730 - 10/18/16 11:56 AM (7 years, 3 months ago) |
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What about pouring off the excess liquid in SAB? too risky?
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azur
God of Fuck



Registered: 04/21/12
Posts: 28,103
Loc: Daid
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Re: TIFU agar soaked in LC [Re: tump]
#23748732 - 10/18/16 11:57 AM (7 years, 3 months ago) |
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Quote:
tump said: Start over again. And let it be if you have the spare jars. You are injecting lc to agar to test the clean of the lc
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blackout


Registered: 07/16/00
Posts: 5,266
Last seen: 2 months, 25 days
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Quote:
mushhiehunter said: What about pouring off the excess liquid in SAB? too risky?
It is risky since the liquid is likely to be leaving nutrients at the top edge of the jar, maybe even spilling over and onto the threads, then contams could grow and track into the jar.
It could be sucked back up, but if it then showed contamination you might be left wondering if contams got in at that time.
Next time you could simply suck up a tiny bit of LC and inject it all, so it can never be too much.
I posted these again just yesterday, tips on syringe control.
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blackout said:
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third3y3 said: It's SOOO easy to squeeze too much. I guess I'll get better. Typical newbie type mistake.
Theres a trick to stop this, do not use the syringe like people typically do. I am right handed, I grasp the syringe in my right hand making a thumbs up sign, the plunger end is sticking up with my thumb under the bit you usually press down with your finger. So if I gave a thumbs up out straight the needle is facing the floor.
My right pinky finger is gripping the body of the syringe at the 10ml sign
 Now you can press down the plunger with your left hand while actively pressing upwards with your right thumb. This gives you good control. If a syringe is unused for a while the rubber seal will be subject to static fritcion (aka stiction). If you use a syringe in a normal manner the moment this stiction is overcome the plunger will tend to move far too much due to the force you are applying and the fact nothing is stopping the syringe from going all the way down.
You can do it with just 1 hand too, just practice with water.
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blackout said:
I didn't state the obvious, but as the syringe empties obviously my pinky finger moves down the body with the print on it. You can get decent control with just 1 hand too, I usually only use the 2 hand method with a syringe which has not been used in a long time. With the 1 hand method I clench my fist around the syringe. When full my little finger wrapped around the syringe body (with the print) and my other 3 fingers grasp the plunger shaft firmly and my thumb presses down. So the plunger shaft is stopped from moving quickly by the friction of my fingers. As the liquid gets used up I put the next finger on the printed body. For the last 2ml my forefinger is just pressed against the underside of the plunger shaft which again just stops it being able to jump down too much.
Another 1 handed method is having your 3 fingers grasp the body with the printing, and your forefinger wrapped around the plunger shaft which provides resistance. Then press down with your thumb in a controlled manner. As said, just practice with plain water and see what suits you best.
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bodhisatta 
Smurf real estate agent


Registered: 04/30/13
Posts: 61,889
Loc: Milky way
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Re: TIFU agar soaked in LC [Re: blackout]
#23748885 - 10/18/16 01:05 PM (7 years, 3 months ago) |
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It would help to plate with the correct technique.
Drop of LC or spores on a loop then streak the plate
Trusted Cultivator said: Doing this dramatically helps your chances
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Trusted Cultivator said:
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Trusted Cultivator said:
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bodhisatta said:
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bodhisatta said:
take your spores to an inoculation loop. if you have a print then flame the loop dip in agar, pick spores off print and then swab the dish
if you have a syringe flame the loop and drop a few drops onto the loop to cool it down and then one more drop to load it with spores and then swab that


Quote:
bodhisatta said: Try using an inoculation loop. Flame the syringe needle and the loop then squirt onto the loop to cool the needle and the loop. Squirt another drop to prime the loop. Then swab the dish.

 
Then you can see growth immediately, see contamination easier, grab healthy mycelium easier, and never worry about spore drops rolling all over a dish giving you germination everywhere at the same time.
also the agar link in my signature
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