exp+15+16+sum08

exp+15+16+sum08 - #12 Endospore stain Green- endospores...

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Unformatted text preview: #12 Endospore stain Green- endospores Red- vegetative cells Bacillus megaterium R E S U L T S a c i d - f a s t b a c t e r i a n o n a c i d - f a s t b a c t e r i a #13 Acid-fast stain Mycobacterium smegmatis Escherichia coli Use of micropipettors (p. 72)- 3 types: 2/20 yellow button. Measures 2-20 μ l 20/200 gold button. Measures 20-200 μ l 100/1000 blue button. Measures 200-1000 μ l • Always use new sterile tip for each transfer. • Discard tips in vesphene beaker 1000 μ l = 1 ml 100 μ l= 0.1 ml Serial dilutions (p. 73) • To count number of microbes present in a sample, must dilute it several times to get a countable number of cells. Dilution = volume transferred volume transferred + volume of blank __1 ___ = __1 ___ 1 + 99 100 • Bacterial numbers are so high that a series of these dilutions is necessary to achieve a countable (30-300; statistically significant) plate. • Purpose- determine number of colony forming units per milliliter (cfu/ml) in a sample. cfu/ml in original culture # of colonies on countable plate reciprocal of the dilution factor of the countable plate X = Example: (52) 100 1 = 5200 = 5.2 x 10 3 cfu/ml S E R I A L D I L U T I O N S O B J E C T I V E : T o d e t e r m i n e t h e n o . o f c o l o n y f o r m i n g u n i t s / m l i n t h e o r i g i n a l c u l t u r e . d i l u t i n g t h e b r o t h c u l t u r e t o o b t a i n a c o u n t a b l e p l a t e c o u n t a b l e p l a t e i s 3 0 - 3 0 0 c o l o n i e s # o f c o l o n i e s o n x r e c i p r o c a l o f t h e d i l u t i o n f a c t o r c o u n t a b l e p l a t e o n t h e c o u n t a b l e p l a t e 1 5 9 x 1 0 3 = 1 . 5 9 x 1 0 5 pp. 73-77 Dilution = volume transferred ....
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This note was uploaded on 10/01/2008 for the course BIOL 2051 taught by Professor Brininstool during the Summer '07 term at LSU.

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exp+15+16+sum08 - #12 Endospore stain Green- endospores...

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