glass beads for plating bacteria to get an even distribution on agar
Pete Muriana
MURIANAP at FOODSCI.PURDUE.EDU
Sat Dec 10 09:57:15 EST 1994
>In article <3c098h$3sc at nuscc.nus.sg>
>phstayhn at leonis.nus.sg (Dr Tay Hou Ngee Agnes) writes:
>> Hi, Does anyone know a supplier and catalog no for glass beads used to
>> spread bacteria in transformation (instead of spreader made from pasteur
>> pipet)? Please reply to phstayhn at leonis.nus.sg
>> Thanks
>> Agnes
Yes, I've used the "glass bead" method since around 1985 when I picked it up
from some friends during my graduate work - it's my preferred method of
surface plating because it spreads the cells so homogeneously on the plate
that you can never get such an even distribution of colonies with a glass
hockey stick (at least I could't :>).
Fisher Sci. sells "Economical solid glass beads" ; we use their 4 mm beads
(#11-312B; $19.00/lb).
For those interested in how to use them:
1. distribute to small test tubes (13mm x 100mm) using a small funnel;
usually 4-6 per tube is sufficient; we usually make one or two racks of
tubes at a time; autoclave them to sterilize.
2. When plating (after sample is aliquoted onto the agar), uncap a tube and
pour the beads onto the plate; stack the plates and slowly shake them
back and forth; every few seconds turn the stack; the idea is to have
the beads travel back and forth along the diameter of the plates, not
the periphery!; usually about 2-3 min. is sufficient; I can do up to 10
plates at a time (I've got big hands :>).
3. When done, tilt a plate on its side, crack it open and pour the beads
into a waste container (preferably containing sanitizer). When you collect
enough "spent" beads, we usually add soap and autoclave them, then rinse
with deionized water, decant excess water, and let them dry for reuse!
This is great for getting evenly-distributed colonies either for plate counts,
colony-lifts, etc. Guaranteed once you try it, you won't plate by any other
method.
-Regards, Peter
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* Peter M. Muriana, Ph.D. 317-494-8284 TEL *
* Dept. of Food Science 317-494-7953 FAX *
* Purdue University murianap at foodsci.purdue.edu *
* Smith Hall *
* W. Lafayette, IN 47907-1160 *
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