NYMU Taipei/Lab Notes/2007 8 6&7

From 2007.igem.org

(Difference between revisions)
Line 14: Line 14:
**** right: <font color="green">GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA</font><font color="red">GGACATGGGTGTGTAGAAGA</font>
**** right: <font color="green">GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA</font><font color="red">GGACATGGGTGTGTAGAAGA</font>
* purchase of BCRC human insulin clone (ordered not paid)
* purchase of BCRC human insulin clone (ordered not paid)
-
* cutting sites of parts
+
* cutting sites of iGEM 2007 parts
** ompR binding ompC promoter
** ompR binding ompC promoter
*** ompR binding site (C1, C2, C3) extraction
*** ompR binding site (C1, C2, C3) extraction

Revision as of 14:38, 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
  • purchase of BCRC human insulin clone (ordered not paid)
  • cutting sites of iGEM 2007 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
  • antibiotics LB plate preparation
    • ampicillin
    • kanamycin