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Team:Cambridge/Protocols/Transformation of B. subtilis
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| - | + | ==Transformation of ''Bacillus subtilis''== | |
| - | + | A technique used to introduce foreign DNA into competent Bacillus cells. | |
| - | == | + | |
| - | + | ||
===Theory=== | ===Theory=== | ||
| - | + | ''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. | |
| + | |||
===Practice=== | ===Practice=== | ||
| - | + | ||
| + | * Add to a clean Eppendorf tube: | ||
| + | <center> | ||
| + | :{| border="1px" style="text-align:center;" | ||
| + | |'''Reagent''' | ||
| + | |scope="col" width="80" | '''Quantity''' | ||
| + | |- | ||
| + | |scope="col" width="200" | competent ''Bacillus'' cells | ||
| + | |400μl | ||
| + | |- | ||
| + | |DNA | ||
| + | |3μl | ||
| + | |- | ||
| + | |} | ||
| + | </center> | ||
| + | |||
| + | * Incubate at 37ºC for 1 hour. | ||
| + | * Plate on an agar plate containing antibiotic, which allows for selection of successful transformants. | ||
| + | |||
===Safety=== | ===Safety=== | ||
| - | + | All equipment (including gloves) that may have come into contact with the bacteria must be autoclaved for decontamination purposes. | |
| - | {{Template:Team:Cambridge/ | + | {{Template:Team:Cambridge/CAM_2011_PROTOCOL_FOOT}} |
Latest revision as of 20:24, 21 September 2011
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Contents |
Transformation of Bacillus subtilis
A technique used to introduce foreign DNA into competent Bacillus cells.
Theory
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.
Practice
- Add to a clean Eppendorf tube:
Reagent Quantity competent Bacillus cells 400μl DNA 3μl
- Incubate at 37ºC for 1 hour.
- Plate on an agar plate containing antibiotic, which allows for selection of successful transformants.
Safety
All equipment (including gloves) that may have come into contact with the bacteria must be autoclaved for decontamination purposes.
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