Transferring Cultures from Slants

I have just rediscovered a few pictures in my website that might be of help to you in the most important aspect ahead of you - preserving the quality of your culture tubes. The fancy word is "axenic", which means 'without any unknowns' - a 'pure culture' with no contaminants.

In this picture you see a person holding an "inoculating loop" (often abbreviated to "loop") in one hand - and a squirt bottle in the other. Know what a "loop" is. I have sent you a bunch of sterile cotton swabs to substitute for a loop.

To use a swab: First, you must realize that the swabs are only sterile inside of their sealed wrappers, so don't think that it is Christmastime and unwrap everything before you use them! Once you have gotten your mind in control of "busy hands", we can proceed.

If you want to know what is inside of each little package, you can feel the sticks and cotton through the paper, but a better observation can be made with "x-ray" vision: hold a packet up to a bright light and you can see two swabs inside.

Now you MUST have a flame (candles don't work - too much sooty flame). (Use a Bunsen burner, and if you don't have one, then use a plumbers' torch - available at your local hardware store. It will come as a set in a box - a small propane tank, a nozzle for adjusting the flame and a sparker to light the thing. The whole works costs about $8.) Light the flame, and then go to the nest paragraph.

The way you get at the swabs is to inspect both ends of the package. One end (but not the other) has two peel-back tabs. This is the end opposite to where the cotton is. Now let's pretend that you are ready to transfer some bacteria from one of the two slants I sent you onto some petri plates. (This implies that you have some agar plates made, dried, and ready to receive the bacteria.) Only then do you peel back the ears about an inch so that you see the two handle-sticks. The first thing you do is touch only the two sticks (and not the peeled back paper) and push the two sticks apart. The reason you do this is so that when you pull one of the swabs out its cotton will not touch the other stick that you just touched.

All the remaining steps much be completed in 5 seconds without any talking.

Step Description Figure, if any
1 Pull out one swab and continue holding it just by the very end of the stick. Set the other one in its package down. Be VERY careful that the cotton touches nothing else except what you want it to touch!
2 Pretending that the large thing in the figure's hand is your swab, use your swab hand's pinkie to unscrew and hold the cap of the slant. (Do NOT put the cap down!)
3 Briefly flame the screw part of the tube to burn away any contaminants. 'Briefly' means a half second. Try always to hold the open tube horizontally to prevent any dust from falling into it.
4 Slip your swab into the tube trying to aim it so well that it touches as little of the opening of the tube as possible. Once you can touch the cotton onto the bacterial scum, give the swab a spin so that all sides touch the bacteria. Then withdraw the loaded swab again trying not to touch the mouth of the tube. (At this time you are holding the following things: the open tube in one hand, and the loaded swab and the tube's cap in the other hand.)
5 Again flame the open end of the tube. This time flame it for about 1 second to heat it more. (Remember that excessive heating may cause the glass to crack! That's a "no-no!")
6 Screw the tube back into the cap that is being held by your pinkie. Don't miss the cap: the tube is hot now and will burn you if you miss! Once the tube is partially treaded into its cap, let go of the cap and have your partner screw the cap on more tightly.
7 Once the tube is set down and out of the way, start using the loaded swab to paint stripes onto your plates. One loaded swab will last for several dozen stripes, so you don't need to reload it.
8 You are now ready to use the other bacterial strain, so repeat the above process using the other swab and the other culture tube.

Step	Description	Figure, if any
1	Pull out one swab and continue holding it just by the very end of the stick. Set the other one in its package down. Be VERY careful that the cotton touches nothing else except what you want it to touch!
2	Pretending that the large thing in the figure's hand is your swab, use your swab hand's pinkie to unscrew and hold the cap of the slant. (Do NOT put the cap down!)
3	Briefly flame the screw part of the tube to burn away any contaminants. 'Briefly' means a half second. Try always to hold the open tube horizontally to prevent any dust from falling into it.
4	Slip your swab into the tube trying to aim it so well that it touches as little of the opening of the tube as possible. Once you can touch the cotton onto the bacterial scum, give the swab a spin so that all sides touch the bacteria. Then withdraw the loaded swab again trying not to touch the mouth of the tube. (At this time you are holding the following things: the open tube in one hand, and the loaded swab and the tube's cap in the other hand.)
5	Again flame the open end of the tube. This time flame it for about 1 second to heat it more. (Remember that excessive heating may cause the glass to crack! That's a "no-no!")
6	Screw the tube back into the cap that is being held by your pinkie. Don't miss the cap: the tube is hot now and will burn you if you miss! Once the tube is partially treaded into its cap, let go of the cap and have your partner screw the cap on more tightly.
7	Once the tube is set down and out of the way, start using the loaded swab to paint stripes onto your plates. One loaded swab will last for several dozen stripes, so you don't need to reload it.
8	You are now ready to use the other bacterial strain, so repeat the above process using the other swab and the other culture tube.

So that's how to do it.