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Offlineozzysmygod
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LC problems. Clumpy not cloudy.
    #14449997 - 05/14/11 08:04 AM (12 years, 9 months ago)

On the 2nd of May I inoculated some Honey LC's with agar wedges. Since then, growth has been slow and the mycelium is forming in a clump around the agar wedge, rather than a cloud in suspension. How can I rectify this?

I tried two types of honey and growth is the same in both types.
I used 10ml of honey to 250ml of water- 4%

Can any UK growers recommend a honey they have had success with? Ive ordered some Karo to see if that improves things.


Thanks

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Invisiblemaug
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14450013 - 05/14/11 08:16 AM (12 years, 9 months ago)

I would guess that when you're ready to pull from it, shake it up really well. I did an LC for my first time too. Mine were also in clumps, and I used a mass spore inoculation. I shook one of the jars for just a couple seconds and it burst into a bunch of small particles (I had a marble in it). I'd assume that shaking it up would also allow it to colonize faster.

I don't think there's really much difference between karo, molasses, etc. I'd just use whatever you have lying around. I happened to have some karo, and it's not like I'm going to be drinking it anytime soon. They are all basically just sugar.


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I think nighttime is dark so you can imagine your fears with less distraction. -Calvin and Hobbes

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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: maug]
    #14450019 - 05/14/11 08:19 AM (12 years, 9 months ago)

Its got a micropore tape filter so I cant shake it too vigourously, to prevent the filter from getting wet.

Edited by ozzysmygod (05/14/11 08:19 AM)

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Invisiblemaug
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14450038 - 05/14/11 08:28 AM (12 years, 9 months ago)

If you're going to use the entire LC in one go, it may not be a problem. If you're going to get it wet from shaking it up anyways, then you might dab a little hydrogen peroxide on it first (up to 10ml, 4%). I'd wait for a second opinion on that though.

If you plan on re-using those lids for LC's, you could make a magnetic stirrer.


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I think nighttime is dark so you can imagine your fears with less distraction. -Calvin and Hobbes

Edited by maug (05/14/11 08:33 AM)

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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: maug]
    #14450074 - 05/14/11 08:42 AM (12 years, 9 months ago)

I dont use hydrogen peroxide. and putting 10ml of Hydrogen Peroxide will probably just make things worse, considering it would more than likely drip through the bringing contaminates with it.

Thanks for your advice, but I was hoping someone with actual LC experiance could help

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OfflineApple core
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14450131 - 05/14/11 09:03 AM (12 years, 9 months ago)

I wouldn't worry too much man. I do a lot of LC work, and used to use a magnetic stirrer. Now I just syringe up the LC then blast it back into the jar to break up the colonies. Never had any problems with this method.

I use Tesco organic honey, it comes in a little squeezy bottle so getting your measurement right is easy. However I use 4% by weight, not by volume. I imagine if you're having problems with growth, there might be too much honey in there.

1 ml of water weighs 1g. But honey is around 36% more dense, so 1 ml will weigh about 1.36 grams. That means you're putting in 13.6g to every 250 ml, which is about 5.4% by weight. So if you want to do it by volume, you should be putting a tad over 7ml into every 250ml of water.

I don't know how much 3 ml is really going to matter, but I'm a bit anal when it comes to precision :nerd:

Hope that helps!

Edited by Apple core (05/14/11 09:04 AM)

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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: Apple core]
    #14450263 - 05/14/11 09:39 AM (12 years, 9 months ago)

Yeah I realised the mistake measuring by volume not weight. Ive started a new batch with 10g of Honey, which I worked out as about 7.5ml. Growth doesnt appear much better though.

At this point I think the clumps are too big to suck up witha syringe. In each jar theres 2-5 big clumps, but no smaller strands. I think sucking it up with a syringe would just draw in the honey water and no mycelium.

Is it Tesco own brand honey you use?

Thanks

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OfflineApple core
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14450568 - 05/14/11 11:01 AM (12 years, 9 months ago)



<- That's it.

When I break up my LC, my first few syringes are just honey-water, but keep blasting the colony with what's in the syringe and eventually it will break up into bits.

What temperature are you incubating at? Also, I just remembered LC need a lot of air exchange, so a magnetic stirrer might be a good project to start. I leave my cultures until I see growth, then stir them like crazy about twice a day. The aim is to create a vortex that draws air into the LC, replenishing the oxygen levels.

Edited by Apple core (05/14/11 11:08 AM)

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Offlinek00laid
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Re: LC problems. Clumpy not cloudy. [Re: Apple core]
    #14450800 - 05/14/11 11:46 AM (12 years, 9 months ago)

if you can work with agar why the fuck are you trying to make a sugar water LC

put that agar wedge into a grain jar instead of a jar of sugar water.


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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: k00laid]
    #14451399 - 05/14/11 01:30 PM (12 years, 9 months ago)

Im incubating at about 75 F. I swirl them everyday but i am looking to get a magnetic stirrer.

And with regards to adding agar wedges to grain jars, i know this is preferable to LC. But using LC its much quicker, and easier to keep sterile. I'll seal spawn bags, PC, inoculate through the bag, tape the hole, then seal with a heat sealer below the inoculation point. Quick and easy and almost no exposure to potential contaminates, unlike opening the bags and innoculating with grain jars.

Thanks

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Offlinek00laid
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14451404 - 05/14/11 01:31 PM (12 years, 9 months ago)

Quote:

ozzysmygod said:
But using LC its much quicker, and easier to keep sterile.




i think unfortunately you will find this not to be true.


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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: k00laid]
    #14451448 - 05/14/11 01:44 PM (12 years, 9 months ago)

How so, do you care to expand? Loads of people on these forums use LC's succesfully.

Have you ever done an experiment to compare jars inoculated with LC side by side with jars inoculated with agar wedges? If not, do you have any other evidence that LC is less effective?

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OfflineSomeFella
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14451546 - 05/14/11 02:04 PM (12 years, 9 months ago)

Quote:

ozzysmygod said:
Im incubating at about 75 F. I swirl them everyday but i am looking to get a magnetic stirrer.

And with regards to adding agar wedges to grain jars, i know this is preferable to LC. But using LC its much quicker, and easier to keep sterile. I'll seal spawn bags, PC, inoculate through the bag, tape the hole, then seal with a heat sealer below the inoculation point. Quick and easy and almost no exposure to potential contaminates, unlike opening the bags and innoculating with grain jars.

Thanks




With proper sterile procedure you can open a bag in a glove box or with a flow hood and perform a G2G at 99%+ success rate...I do it all the time....

How is an lc where you cannot see contams easier to keep sterile than a petri dish?....one of the main reason people use agar is to clean up cultures.....also....its NOWHERE near as fast as expanding via G2G....I can turn a whole room into fully colonized spawn jars from just one agar plate before you even get yer LC into a syringe...and I will know that my culture is not contamintaed..you on the other hand will want to check yer LC on at least one jar first before you risk knocking up several jars with a contaminated solution....taking up another week or so....you will never convince any consistently successful grower producing considerable amounts that the LCs of today are more sterile or faster than starting with agar and expanding via G2G....its silliness....

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Offlinek00laid
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14451548 - 05/14/11 02:05 PM (12 years, 9 months ago)

depends what you mean by effective.

but it is in fact harder to get a clean jar of mycelium laden sugar water than it is to get a clean jar of grains.


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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: k00laid]
    #14451574 - 05/14/11 02:11 PM (12 years, 9 months ago)

I'll try both methods then, thanks

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OfflineSomeFella
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14451588 - 05/14/11 02:14 PM (12 years, 9 months ago)

Quote:

ozzysmygod said:
How so, do you care to expand? Loads of people on these forums use LC's succesfully.

Have you ever done an experiment to compare jars inoculated with LC side by side with jars inoculated with agar wedges? If not, do you have any other evidence that LC is less effective?




I actually have done several side by sides with different LCs.....and the results are ALWAYS the same....the mycelium from the G2G recovers and gets going much more quickly than any LC I have worked with...

and like I said earlier if you really wanted to race fair you would start by droppin some spores into yer LC the same day you drop spores on an agar plate....you will be waiting for your LC to colonize while im already expanding grains...also gotta mention the LC test jar...right?...cause we all know thats what you should do if you were serious at all about success with LCs....and with that extra week added on...the agar and G2G method easily trumps LC.....easily.....on all fronts....

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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: SomeFella]
    #14451605 - 05/14/11 02:17 PM (12 years, 9 months ago)

Okay, thanks for the info. I'll inoculate some more plates and use them for inoculation and G2G. How big a sized wedges do you use to inoculate quart jars?

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Offlineozzysmygod
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Re: LC problems. Clumpy not cloudy. [Re: ozzysmygod]
    #14456791 - 05/15/11 03:07 PM (12 years, 9 months ago)

Bump

What sized wedges do people inoculate quart jars with? Thanks

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