NYMU Taipei/Lab Notes/2007 8 6&7
From 2007.igem.org
Lab schedule
- mouse insulin gene cloning (delay to 8/7)
- primer design for insulin II of Mus musculus, MN_008387.3 transcript
- 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 for A chain
- 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
- tetR-EnvZ fusion construct
- antibiotics LB plate preparation
- ampicillin
- kanamycin