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| - | ==Transformation of ''Bacillus subtilis''==
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| - | A technique used to introduce foreign DNA into competent Bacillus cells.
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| - | ===Theory===
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| - | ''Bacillus subtilis'' is a rod-shaped, soil-dwelling bacterium . It is widely used as a model organism for studying biology of Gram-positive bacteria which, in contrast to Gram-negative ''E.coli'', do not possess the outer membrane over the peptidoglycan cell wall. Additionally, ''Bacillus'' is naturally competent, which means that no sophisticated pre-treatment is needed for transformation with foreign DNA.
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| - | ===Practice===
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| - | * Add to a clean Eppendorf tube:
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| - | <center>
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| - | :{| border="1px" style="text-align:center;"
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| - | |'''Reagent'''
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| - | |scope="col" width="80" | '''Quantity'''
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| - | |-
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| - | |scope="col" width="200" | competent ''Bacillus'' cells
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| - | |400μl
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| - | |DNA
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| - | |3μl
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| - | |}
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| - | </center>
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| - | * Incubate at 37ºC for 1 hour.
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| - | * Plate on an agar plate containing antibiotic, which allows for selection of successful transformants.
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| - | ===Safety===
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| - | All equipment (including gloves) that may have come into contact with the bacteria must be autoclaved for decontamination purposes.
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| - | <center>'''[[Team:Cambridge/Protocols|Back to Protocols]]'''</center>
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Latest revision as of 15:25, 13 July 2012
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