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 Chk
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Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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 Simple way to multiply LI without introducing contaminants 
#23422382 - 07/08/16 04:45 AM (8 years, 3 months ago) |
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For the record i was using the simple way to make LI : scraping a plate with the needle and sucking everything in.
Also i'm kind of lazy and i often forget stuff around ...
I realized you always scrape a bit of agar when you're making LI that way, which end up melting in the water of the syringe.
Briefly : i ended up with a liquid culture coming from a LI syringe just by shear lazyness XD
So here is my VERY simple proposal:
1 - Take a clean plate to make your LI
2 - Scrape the surface hard with your needle (the point being to scrap some more agar than usual here)
3 - Suck back everything in your syringe
4 - just wait for the mycélium to develop again
5 - you just multiplied your LI by a shitload without chance for contams to enter
Obviously it's better to use a bigger syringe here !
And also not a too tiny needle which could end up clogged by the mycélium.
That syringe was containing only one or two minuscule strands of mycélium,
it ended up fully colonized, and i shaked it a bit so we can see better:
Edit: i have some 50ml syringe left, i'm gonna test with those next.
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Edited by Chk (07/08/16 04:48 AM)
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 Munchauzen
Registered: 06/22/11
Posts: 14,348
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 Re: Simple way to multiply LI without introducing contaminants [Re: Chk]
#23422530 - 07/08/16 06:45 AM (8 years, 3 months ago) |
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sounds complicated
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Supalemonhaze
Spore syringe hater.
Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
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Re: Simple way to multiply LI without introducing contaminants [Re: Munchauzen]
#23422594 - 07/08/16 07:20 AM (8 years, 3 months ago) |
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The amount you gain by letting it recover is not worth the trouble it's gonna cause by clogging the needle. Just scrape the mycelium and use it instantly if you need it, a colonized agar plate is easy enough to get. If more mycelium is what you are after, inoculate an extra plate instead of doing this and you will end up having more anyway.
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bodhisatta  
Smurf real estate agent
Registered: 04/30/13
Posts: 61,901
Loc: Milky way
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Re: Simple way to multiply LI without introducing contaminants [Re: Supalemonhaze]
#23422617 - 07/08/16 07:30 AM (8 years, 3 months ago) |
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Scraping plates has got to be the worst way to make the LI too. Blender or blenderless LI is superior the plate scrape works in a pinch but at that point I just take wedges and use those.
Any LI will recover if left to sit. The purpose of LI is immediate use. The mycelium for the LI has to be growing on something so when you make LI and let it sit you just made bootleg LC
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Chk
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Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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Re: Simple way to multiply LI without introducing contaminants [Re: Chk]
#23422773 - 07/08/16 08:44 AM (8 years, 3 months ago) |
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Well i don't know for you but i don't have hundreds of petris around !
most are plates in the fridge that i keep for further use.
I also disagree about it being complicated since it's exactly the same stuff as making a LI with your method ,munchauzen (which i like a lot)
Basically what i'm proposing is a way to make a LC without the problem usually involved.
Given that you use a big syringue it is a lot safer than using your plate to make a LC.
But i can agree it's maybe not the most time efficient method.
Not an issue for me.
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Edited by Chk (07/08/16 08:46 AM)
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Chk
Reverser
Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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Re: Simple way to multiply LI without introducing contaminants [Re: Supalemonhaze]
#23422780 - 07/08/16 08:47 AM (8 years, 3 months ago) |
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Btw i'm using 14 gauge needles, clogging isn't really an issue for me either
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Supalemonhaze
Spore syringe hater.
Registered: 10/02/15
Posts: 6,725
Loc: 12" down Europe's butthole
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Re: Simple way to multiply LI without introducing contaminants [Re: Chk]
#23422800 - 07/08/16 08:54 AM (8 years, 3 months ago) |
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Still, you are much better off making extra plates and using those instead. If you can't afford petris (which is kinda ridiculous since disposables are cheap enough), you can use pp5 containers which are re-usable forever.
If you are using ports they will last significantly less with a 14g needle than a 16g one (hence why 16g is recommended). Seems to me that for a negligible amount of inoculant, you are increasing risk, increasing maintenance of your jars as well as increasing the time you have to wait until inoculation. You seem to be focusing on all of the wrong things. IMO, spending 5cents extra for a petri is infinitely better than having to replace your ports twice as often, but that's just me. To each his own I guess.
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mupetmower
Mower of Muppets
Registered: 03/29/16
Posts: 3,036
Loc: Here and There
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Re: Simple way to multiply LI without introducing contaminants [Re: Chk]
#23422810 - 07/08/16 09:00 AM (8 years, 3 months ago) |
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Yeah I completely disagree. This is much more complicated than Munch's blenderless LI and it also introduces MUCH MORE potential bacteria/contam vectors. I hardly fail to see the benefit of doing this.
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-The wise man never stops seeking knowledge.
   
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
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MycoLoopology
Eye's wide open
Registered: 06/12/16
Posts: 518
Last seen: 7 months, 2 days
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Re: Simple way to multiply LI without introducing contaminants *DELETED* [Re: Supalemonhaze]
#23422813 - 07/08/16 09:01 AM (8 years, 3 months ago) |
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Post deleted by MycoLoopology
Reason for deletion: privacy
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Chk
Reverser
Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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Re: Simple way to multiply LI without introducing contaminants [Re: MycoLoopology]
#23422821 - 07/08/16 09:07 AM (8 years, 3 months ago) |
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Well , one of you has to come with real argument as to WHY it would increase chance of contams
since i'm directly taking mycelium from the plate into the siryngue.
Also i don't use disposable petris, only glass ones.
Only things i use that are disposable are some siryngue (but still i have many reusable glass ones)
I can only agree on the fact that 14 gauge needles tend to destroy ports :/
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Edited by Chk (07/08/16 09:07 AM)
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Munchauzen
Registered: 06/22/11
Posts: 14,348
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Re: Simple way to multiply LI without introducing contaminants [Re: mupetmower]
#23422844 - 07/08/16 09:15 AM (8 years, 3 months ago) |
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Quote:
mupetmower said:
Yeah I completely disagree. This is much more complicated than Munch's blenderless LI and it also introduces MUCH MORE potential bacteria/contam vectors. I hardly fail to see the benefit of doing this.
I don't know what exact tek OP is using, but I used to have a SCRAPE-LI tek published to my name. I have sense removed it and replaced it with the blenderless because I saw no upside to the scrape anymore.
It's more prone to contamination because you are making direct contact with the culture and the SHIP. Direct contact is always a vector, even more so with poor technique.
Also the agar is colonized itself, so you are reducing inoculation points by not using the agar itself.
Edited by Munchauzen (07/08/16 09:17 AM)
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mupetmower
Mower of Muppets
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Posts: 3,036
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Re: Simple way to multiply LI without introducing contaminants [Re: Munchauzen]
#23422858 - 07/08/16 09:18 AM (8 years, 3 months ago) |
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Yeah more contact = more vectors.
It's simple logic.
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-The wise man never stops seeking knowledge.
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
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Chk
Reverser
Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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Re: Simple way to multiply LI without introducing contaminants [Re: Munchauzen]
#23422862 - 07/08/16 09:19 AM (8 years, 3 months ago) |
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Btw i didn't see your new blenderless LI post.
Note that one could use the same principle i'm doing here with your "new" method.
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Edited by Chk (07/08/16 09:19 AM)
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mupetmower
Mower of Muppets
Registered: 03/29/16
Posts: 3,036
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Re: Simple way to multiply LI without introducing contaminants [Re: Chk]
#23422867 - 07/08/16 09:20 AM (8 years, 3 months ago) |
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Sure, you could, but why the hell would you? That literally makes ZERO sense.
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-The wise man never stops seeking knowledge.
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
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Chk
Reverser
Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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Re: Simple way to multiply LI without introducing contaminants [Re: mupetmower]
#23422884 - 07/08/16 09:24 AM (8 years, 3 months ago) |
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well i don't inoculate jars every day, i could make some LI , just let it sit, and have a far more dense LI just day afters.
So, maybe it's not useful for everyone that's granted, but saying it make no sense just astonish me.
Less work = make sense
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Edited by Chk (07/08/16 09:24 AM)
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mupetmower
Mower of Muppets
Registered: 03/29/16
Posts: 3,036
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Re: Simple way to multiply LI without introducing contaminants [Re: Chk]
#23422889 - 07/08/16 09:25 AM (8 years, 3 months ago) |
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But... How are you getting less work out of this? What you are suggesting is more work.
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-The wise man never stops seeking knowledge.
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
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Chk
Reverser
Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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Re: Simple way to multiply LI without introducing contaminants [Re: mupetmower]
#23422899 - 07/08/16 09:28 AM (8 years, 3 months ago) |
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i'm suggesting that by having a more dense inoculant i can use less.
or eventually that i can use the same quantity and get better result (unsure)
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Edited by Chk (07/08/16 09:29 AM)
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MycoLoopology
Eye's wide open
Registered: 06/12/16
Posts: 518
Last seen: 7 months, 2 days
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Re: Simple way to multiply LI without introducing contaminants *DELETED* [Re: Chk]
#23422913 - 07/08/16 09:32 AM (8 years, 3 months ago) |
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Post deleted by MycoLoopology
Reason for deletion: privacy
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Chk
Reverser
Registered: 09/06/13
Posts: 405
Loc: France
Last seen: 6 years, 13 days
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Re: Simple way to multiply LI without introducing contaminants [Re: MycoLoopology]
#23422926 - 07/08/16 09:36 AM (8 years, 3 months ago) |
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i guess i should have also said that my SAB is not very big, and that i can't take a bigger one right now, (i'm very very short on space to put things)
that limits me quite a lot in what i can do (G2G transfer are nearly impossible in my SAB for example)
Hopefully i should be able to move soon, and have a dedicated room for growing 
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Edited by Chk (07/08/16 09:37 AM)
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mupetmower
Mower of Muppets
Registered: 03/29/16
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Re: Simple way to multiply LI without introducing contaminants [Re: MycoLoopology]
#23422927 - 07/08/16 09:36 AM (8 years, 3 months ago) |
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Also, having a "denser" LI does not equate to less work, even if you are suggesting as such for the long run. Even if we stick with this "denser" LI thought, wouldn't just dripping a whole plate into your sterile water give a "denser" LI than scrapping only a part of it in? So less work for your "denser" LI..
And if your SAB is big enough to for a quart, hell even a pint jar in, which I can't see there being any SAB that small, then you can do blenderless LI..
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-The wise man never stops seeking knowledge.
-I wanna feel the change consume me, feel the outside turning in. I wanna feel the metamorphosis and cleansing I've endured within.
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