Question - liquid innoculation

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By Ryan Waters (Zerogravity) on Monday, December 17, 2001 - 12:33 am:

How long before I can see growth from my liquid innouculation?

Here is what I have done. First, I followed all tek procedures that I have read. This included autoclaving everything, HEPA filtering, sterilizing a clean room, innouculating the jars when they were at room temp, gloves, & lab suit. I used poly fiber fill as a lid vent filter underneath a tin foil cover. I also placed 2 sterilized nickels in the jar for agitation purposes.

I mixed in a 1.5 litre jar, 1 litre of sterilized reverse osmosis water, 4% (by volume) of Crown corn syrup (could not find Karo light corn syrup as read in this forum), and 10 cc of B+ spores in a commercial syringe. The ingredients of the syrup are as follows. Glucose, glucose-fructose, water, salt and vanillin. Question do you know what vanillin is?

I then placed the jar in a dark and stable cabinet at 86 degrees for 72 hours. This room is HEPA filtered.

At this point in time the jars are beautifully crystal clear but of course I want to see anything but this...I want growth! Do you know when can I expect to see something? (I thought I read that I would see something within 48 - 72 hours)

My thanks in advance.

By quote: (Quote) on Monday, December 17, 2001 - 12:40 am:

that's a bit optimistic, but not unreasonable.
3-7 days is the range, so you still could/should see something soon.

By Ryan Waters (Zerogravity) on Monday, December 17, 2001 - 03:23 am:

Thanks for the quick response Quote!

I forgot to mention that the 2 nickels in the jars have turned all rusted / green & brown. I scrubbed them vigorously with alcohol, then let them soak in alcohol for one minute, rinsed with tap water, then autoclaved them.

Pls tell me I'm OK :)

By quote: (Quote) on Monday, December 17, 2001 - 04:06 am:

nickels in the jar ?
hmm... i use bits of broken glass.
wonder if something toxic leached from your nickels...

By Brad (Raze) on Monday, December 17, 2001 - 04:37 am:

I'd use a rock before I'd use metal in there

By Nan (Nanook) on Monday, December 17, 2001 - 06:49 am:

Break a jar and put some glass fragments in the liquid culture jar if you feel you need some maceraction action before drawing up syringes.

Be careful and do not allow the polyfil to get wet by tipping or shaking the jar. Really, sealed jars work much better: lower contam rate and you can shake them to your hearts content without leakage.

I am going to remove the reference to the nickels from the archive. I see problems with that already. I had one drop on the shower floor and sit for a few days and they do corrode some.

By Imok Urok2 (Imok) on Monday, December 17, 2001 - 06:55 am:

Since 1965 nickels are not just made up of silver.
They are a sandwich of silver with a metal alloy center.
I don't know about a rock(what type of mineral?), but couldn't the
shaking, with them bouncing around inside, break the jar?
Sterilized glass(silica) sounds like a better bet :)
Just thought of this.
Like Ensign Pulver, how about one sterilized glass marble?
It would be easy to use it to agitate the liquid, just roll it around the jar.
Never tried it, just a suggestion :)

By Ryan Waters (Zerogravity) on Monday, December 17, 2001 - 11:54 pm:

Thanks for all the input guys...I will take your advice. Hope I can return the favor sometime.

I will wait to see if the jars develop growth but just in case I will start fresh. This time I will use sealed jars as per Nan and use marbles instead of nickels. I will keep you informed if something does develop in the jars with the nickels.

Question: What is Vanillin in the corn syrup? Why does the tek suggest the use of lite as opposed to regular corn syrup? Will this make a difference? I ask cause after two stores I could not find the lite stuff. Of course, I will keep searching if it is better.

By Brad (Raze) on Tuesday, December 18, 2001 - 12:30 am:

I only meant a small rock, not a big huge rock.
If in doubt, put something in water for a few days to make sure it doesnt rust on you

By Ryan Waters (Zerogravity) on Tuesday, December 18, 2001 - 02:14 am:

Yes I've got more questions than I have patience but I'm real concerned about doing things right.

In the liquid inoculation, do I add 4% of corn syrup by weight or volume?

By quote: (Quote) on Tuesday, December 18, 2001 - 02:28 pm:


By Ryan Waters (Zerogravity) on Tuesday, December 18, 2001 - 07:52 pm:

I was unable to find light corn syrup in the two stores I went to but did of course find regular. Has anyone tried both? Would the regular be better or worse?
(I looked in the archives for this answer prior to posting and must have missed the answer if it was there.)


By Nan (Nanook) on Tuesday, December 18, 2001 - 10:00 pm:

The reason Light Corn Syrup was specified was for clarity. I imagine the regular corn syrup would work, but it is darker and the Tek jar will have some tint as a result.

I determined the actual dry sugar weight of one teaspoon Light Corn Syrup as being 4 grams so you can use a liquid measure (you don't have to weigh). If you do weigh syrup, remember you are weighing sugar & water, not pure sugar. The only way to know how many grams of sugar are in a gram of Corn Syrup is to weigh a sample, desiccate it, and weigh the sugar remaining... Trust me, this uses common kitchen measures after I worked with several samples

If you need larger batches just add one additional teaspoon of syrup for each additional 100 ml of water. ml = cc you can use an empty syringe to measure.

By Ryan Waters (Zerogravity) on Wednesday, December 19, 2001 - 12:31 am:

Are we talking light in color or light in sugar content?

We used the syrup that was light in color for clarity purposes but if there is a syrup that is light in sugar that you are using then this will definetly affect things.

Thanks for the advice on the weights guys

By Nan (Nanook) on Wednesday, December 19, 2001 - 07:42 am:

Light as in color. Sugar content as in standard Corn Syrup. You can clearly read a typical product label in one of the photo's in the TEK. I have purchased several brands, all with labeling close enough to the posted photo that there should little chance of error.

This is a standard grocery item long used in the making of sweets and desserts. It's nothing fancy, new, or improved. Usually found in the pancake syrup section.

By Tom Baxter (Kappy) on Wednesday, December 19, 2001 - 08:57 pm:

can someone tell me how much malt powder to use in a half pint jar for liwuid innoc?

By quote: (Quote) on Thursday, December 20, 2001 - 06:12 pm:

4% by weight, that is, 4 grams for every 100 cc water.