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Say hello to my little friend Reged: 09/15/12 Posts: 37808 Loc: Canada |
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Pastywhyte's EzLC Like many people I am a closet LC enthusiast. I find it great to inoculate bags and the speed is nice as well. But of course LC is a method that carries high contam vectors and needs a little extra care. With my EzLC we can visually inspect the culture before we introduce the liquid meaning as long as your confident in the liquid phase you can be sure of a clean LC (test it at first to be sure you have your movements etc down). We also are able to make LC as easy as transferring agar wedges, with the same action as blenderless, but an aggressive colonizing culture with plenty of stored energy to recover. I find that with this LC I often see recovery in 48 hours with fast aggressive growth. This EzLC is as simple as making no pour agar and requires little in the way of special equipment. Super EzLC doesn't even require the use of Whatman filters or SHIP's. This is to LC what blenderless LI is to LI. The best part of the method is the elimination of sediment. You can make brf LC. Or potato flakes. All the sediment is trapped in the agar puck. These more complex media's result in quicker recovery after inoculation. I always found karo or SITR to make for sluggish recovery. I have seen 3 day recovery and explosive growth consistently with some of these recipes you normally would avoid for LC. Materials needed: 2 half pint or pint jars. Fancy Jelly Jars also work well. 2 LC lid with whatman and SHIP 1 60cc Syringe. Smaller will work but more will be needed. 1 PC You will also need agar supplies. I like powdered food grade agar and really any nutes that you like works for this. PDA, MEA, GW, DFA are all good and kick ass for this. So the first thing you need to do is mix up some agar. First you need to decide how much LC you plan to make. 120 ml is a nice amount, more than enough to knock up a pile of grains or bags, and if you do this in widemouth half pints, 120 cc fills it pretty good. So for 120 ml final LC I want to be using about 2-4 grams worth of nutes, be it potato or malt etc. I then make up about 50 ml of water with 2-4 grams nutrition and 1 gram agar. This puts the agar at 2% and nutrient rich. I have tailored the above recipe for use of one LC. You will probably want more than that, especially if you plan to use this to test cultures or get bored of growing the same thing all the time. At the end of the day I wouldn't stress to much over ratios etc. Make up some agar, use lots of nutes, make up the jar. Quick example: Bam! Its that damn easy. So now ya pressure cook it up. Once PCd and cool we can now knock these up with a clean culture. Take a clean wedge and put it too the plate same as you would make transfers. Let it grow out to the size of a quarter or so. Do to the high nutes it may grow a touch slower but that's okay cause we don't need it to grow out much. Once I have a clean culture I can visually inspect it for mold or bacteria or other contams. Once I am happy that its clean I then sterilize a jar of water with a lid outfitted with a SHIP. Have a sterilized syringe ready and then using proper sterile procedure aspirate the water and inject into the culture jar. Sterile water and syringe: You will soon notice that the nutes from the agar puck leach into the water (as will the food coloring if you use it) but any sediment or solid matter remain trapped in the agar puck. Give it some time to colonize, usually will take a week to 10 days. Swirling it aggressively will help expand the colony faster. You might dislodge the agar but that is not an issue. Pic of colonized EzLC Once colonized you are free to aspirate and use. It makes for a pretty aggressive and fast recovering LC compared to the simple sugar recipes many people make. BRF LC is damn fast to recover and dog food agar usually provides explosive growth. Grain water, potato and of course malt extract all kick ass. It can get very thick if you let it go to long so be careful, I have had it get pretty solid. Makes it hard to aspirate. You can use any size jar for this but I find a quart to end up with more LC than I. Half pints and 120 ml LC is enough to do 60 quarts easy. More than enough. I also like the fancy Jelly jars. They are so low that often you can aspirate the liquid without tilting the jar. Makes things much easier. Grain water EzLC. Dark but clear. I may test this one because IME media like grain water or dog food can become susceptible to colonies blooming over time. I would never use grain water or dog food recipes for long term use due to high populations inherent to that media. But it looks very good at first glance. Super Ez LC Supposing you want to try this but don't have any LC lids. With taller jars its perfectly safe to use a grain jar lid with SFD and just pour the water and then the LC. You just need to be careful when swirling that you don't get the liquid up to the filter. Taller jar helps with this. Materials needed 2 Grain jar lids with SFD 2 pint jars (I prefer reg mouth but widemouth can work too) Agar supplies. Make your no pour agar the same as with the EzLC. Pour in pint jars and PC. Transfer the wedge as normal but the drop might cause it to bounce off to the side. It doesn't matter if its not dead center, just leave it. When sufficiently colonized sterilize a jar of water and in a SAB or in front of flow hood pour the sterilized water into the colonized no pour jar. As long as the level is kept low (less than 250 ml) its easy to swirl and keep the liquid from hitting the filter. Potato flakes and karo with simple grain jar lids. Note how thick and healthy the colony looks. These were also mostly ignored, very little agitation was done. The jars above are crystal clear and look strong. They could be tested if you had any concerns and I recommend testing or simply tossing anything that looks off but, these I personally am confident would be fine. Once colonized you can simply pour it into your grain jars same as G2G. The colony with this method recovers really fast. Buy a dozen reg mouth pints and keep them rolling real easy. Fast easy spawn.
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