NYMU Taipei/Lab Notes/2007 8 6&7
From 2007.igem.org
(Difference between revisions)
Line 16: | Line 16: | ||
* cutting sites of parts | * cutting sites of parts | ||
** ompR binding ompC promoter | ** ompR binding ompC promoter | ||
+ | *** ompR binding site (C1, C2, C3) extraction | ||
+ | *** assembly to insulin | ||
** [http://partsregistry.org/Part:BBa_C0082 tar-EnvZ fusion construct]: cut EnvZ part for our fusion construct | ** [http://partsregistry.org/Part:BBa_C0082 tar-EnvZ fusion construct]: cut EnvZ part for our fusion construct | ||
* antibiotics LB plate preparation | * antibiotics LB plate preparation | ||
** ampicillin | ** ampicillin | ||
** kanamycin | ** kanamycin |
Revision as of 14:37, 6 August 2007
Lab schedule
- mouse insulin gene cloning (delay to 8/7)
- primer design for insulin II of Mus musculus, MN_008387.3 transcript
- sequence for standard assembly (marked in green) and PCR reaction (marked in red)
- primer for A chain
- translation (90-110): GIVDQCCTSICSLYQLENYCN
- transcript (341-403): GGCATTGTAGATCAGTGCTGCACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAAC, 63 bp
- left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATGATTGTAGATCAGTGCTGCAC
- right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGTTGCAGTAGTTCTCCAGCT
- primer for B chain
- translation (25-54): FVKQHLCGSHLVEALYLVCGERGFFYTPMS
- transcript (146-235):TTTGTCAAGCAGCACCTTTGTGGTTCCCACCTGGTGGAGGCTCTCTACCTGGTGTGTGGGGAGCGTGGCTTCTTCTACACACCCATGTCC, 90 bp
- left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTTTGTCAAGCAGCACCTTTG
- right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGGACATGGGTGTGTAGAAGA
- primer design for insulin II of Mus musculus, MN_008387.3 transcript
- purchase of BCRC human insulin clone (ordered not paid)
- cutting sites of parts
- ompR binding ompC promoter
- ompR binding site (C1, C2, C3) extraction
- assembly to insulin
- tar-EnvZ fusion construct: cut EnvZ part for our fusion construct
- ompR binding ompC promoter
- antibiotics LB plate preparation
- ampicillin
- kanamycin