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| ===Materials===
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| * 1mL sterile syringe (BD 1mL tuberculin slip tip)
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| * 5μm filter (Millex-SV, fits onto end of syringe)
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| ===Directions===
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| # Take a culture of ''M. smegmatis'' in exponential phase. Pipette 1mL into each of two Eppendorf tubes.
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| # Centrifuge the tubes for 5 minutes at 5,000rpm.
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| # Pipette off the supernatant, and resuspend in 100μL 7H9 medium.
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| # Place a clean Eppendorf in a rack. Put the filter on the end of the syringe, and remove plunger from syringe. Place it upside down in another slot of the rack.
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| # Load the contents of both Eppendorf tubes into the syringe with a pipette, replace the plunger, and use it to force the liquid through the filter into the clean Eppendorf. The cells are ready for microscopy.
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| ===Notes===
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| Mycobacteria clump horribly, so to do microscopy sensibly, you have to break up or remove these clumps. All of this should be done in a hood, as with any other manipulation of such slow growing bacteria.
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