User:Isis Trenchard/Notebook/BioE44 Stuff/2010/03/24: Difference between revisions
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This will be held under a repressible promoter on the ''pJexpress cloning cassette from DNA2.0''. We codon optimised the operon for both E. coli and B. subtilis, and designed it to include restriction sites with complementary sticky ends around vioD and vioC. This allowed us to remove both genes easily to create more colours from the vio operon. | This will be held under a repressible promoter on the ''pJexpress cloning cassette from DNA2.0''. We codon optimised the operon for both E. coli and B. subtilis, and designed it to include restriction sites with complementary sticky ends around vioD and vioC. This allowed us to remove both genes easily to create more colours from the vio operon. | ||
==Plan== | |||
3/24 | |||
*transform | |||
3/25 | |||
*inoculate positive colonies | |||
3/26 | |||
*miniprep | |||
*make freezer stocks | |||
*digest | |||
*gel extract | |||
3/28 | |||
*ligate and transform 002, 003, 004 into pSB1A3 | |||
3/29 | |||
*inoculate colonies for freezer stocks | |||
3/30 | |||
*make freezer stocks | |||
Revision as of 17:57, 24 March 2010
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Details of the first part of the classDay 2 (Thurs April 1)
Day 2.5 (Fri April 2, TAs)
Day 3 (Tues April 6)
Day 3.5 (Wed April 7, TAs)
Day 4 (Thurs April 8)
Day 4.5 (Fri April 9)
Day 4.5 (Mon April 12 - students)
Day 5 (Tues April 13)
Other stuff left to order
Pigment extraction protocol
Biobrick plasmids and promotors
Vio operon details
DNA2.0 very generously agreed to synthesize the entire operon for us, we designed it to include all the five genes, each preceded by a ribosome binding site, and flanked by the prefix and suffix. This will be held under a repressible promoter on the pJexpress cloning cassette from DNA2.0. We codon optimised the operon for both E. coli and B. subtilis, and designed it to include restriction sites with complementary sticky ends around vioD and vioC. This allowed us to remove both genes easily to create more colours from the vio operon. Plan3/24
3/25
3/26
3/28
3/29
3/30
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