Imperial College/19 August 2008
From 2008.igem.org
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==Wetlab== | ==Wetlab== | ||
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+ | ===Cloning=== | ||
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+ | *GFP-3-mutB - Terminator and CAT containing XL1-BLues replica plated and overnight culture prepared | ||
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+ | *Terminator, pSB1AK3 and mRFP - Terminator miniprepped from overnight cultures (all 5ml used) | ||
+ | **Single digests (''EcoRI'') run on a gel to determine approximate size | ||
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===''B.subtilis''=== | ===''B.subtilis''=== | ||
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*Transformation protocol 2 was repeated today, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol]. The basic principle of this protocol is that competent cells are prepared by growing a culture to a high O.D.<sub>600</sub> and then performing electroporation on these competent cells. | *Transformation protocol 2 was repeated today, [http://openwetware.org/wiki/IGEM:IMPERIAL/2008/Prototype/Wetlab/Transformation_protocol_4#Transformation_Protocol_2| Click link here for protocol]. The basic principle of this protocol is that competent cells are prepared by growing a culture to a high O.D.<sub>600</sub> and then performing electroporation on these competent cells. | ||
*In previous attempts the competent cells were grown to an O.D.<sub>600</sub> of 1.5. Although this transformation worked, we wished to try to increase the O.D.<sub>600</sub> to increase the efficiency of transformation. The culture was grown to an O.D.<sub>600</sub> of 1.8 before being harvested. | *In previous attempts the competent cells were grown to an O.D.<sub>600</sub> of 1.5. Although this transformation worked, we wished to try to increase the O.D.<sub>600</sub> to increase the efficiency of transformation. The culture was grown to an O.D.<sub>600</sub> of 1.8 before being harvested. |
Revision as of 19:06, 25 August 2008
19 August 2008WetlabCloning
B.subtilis
Miscellaneous
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