NYMU Taipei/Lab Notes/2007 8 6&7

From 2007.igem.org

(Difference between revisions)
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* mouse insulin gene cloning (delay to 8/7)
* mouse insulin gene cloning (delay to 8/7)
** primer design for [http://www.informatics.jax.org/searches/accession_report.cgi?id=MGI%3A96573 insulin II of Mus musculus], [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?cmd=Retrieve&db=Nucleotide&list_uids=142350788&dopt=GenBank&WebEnv=04iJq7mV5tF1N_LSoJWAy8uc-0i_vXvfkGc3KMxcxnrIPG9K-LDVOuYkZWjzh6BpujGcDGNGQk0k9I%40D4770B146B72B990_0041SID&WebEnvRq=1 MN_008387.3 transcript]
** primer design for [http://www.informatics.jax.org/searches/accession_report.cgi?id=MGI%3A96573 insulin II of Mus musculus], [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?cmd=Retrieve&db=Nucleotide&list_uids=142350788&dopt=GenBank&WebEnv=04iJq7mV5tF1N_LSoJWAy8uc-0i_vXvfkGc3KMxcxnrIPG9K-LDVOuYkZWjzh6BpujGcDGNGQk0k9I%40D4770B146B72B990_0041SID&WebEnvRq=1 MN_008387.3 transcript]
 +
*** sequence for <font color="green">standard assembly (marked in green)</font> and <font color="red">PCR reaction (marked in red)</font>
*** primer for A chain
*** primer for A chain
**** translation (90-110): GIVDQCCTSICSLYQLENYCN
**** translation (90-110): GIVDQCCTSICSLYQLENYCN
**** transcript (341-403): GGCATTGTAGATCAGTGCTGCACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAAC, 63 bp
**** transcript (341-403): GGCATTGTAGATCAGTGCTGCACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAAC, 63 bp
-
**** left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATG<font color="red">ATTGTAGATCAGTGCTGCAC</font>
+
**** left: <font color="green">GTTTCTTCGAATTCGCGGCCGCTTCTAGATG</font><font color="red">ATTGTAGATCAGTGCTGCAC</font>
-
**** right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA<font color="red">GTTGCAGTAGTTCTCCAGCT</font>
+
**** right: <font color="green">GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA</font><font color="red">GTTGCAGTAGTTCTCCAGCT</font>
*** primer for B chain
*** primer for B chain
**** translation (25-54): FVKQHLCGSHLVEALYLVCGERGFFYTPMS
**** translation (25-54): FVKQHLCGSHLVEALYLVCGERGFFYTPMS
**** transcript (146-235):TTTGTCAAGCAGCACCTTTGTGGTTCCCACCTGGTGGAGGCTCTCTACCTGGTGTGTGGGGAGCGTGGCTTCTTCTACACACCCATGTCC, 90 bp
**** transcript (146-235):TTTGTCAAGCAGCACCTTTGTGGTTCCCACCTGGTGGAGGCTCTCTACCTGGTGTGTGGGGAGCGTGGCTTCTTCTACACACCCATGTCC, 90 bp
-
**** left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATG<font color="red">TTTGTCAAGCAGCACCTTTG</font>
+
**** left: <font color="green">GTTTCTTCGAATTCGCGGCCGCTTCTAGATG</font><font color="red">TTTGTCAAGCAGCACCTTTG</font>
-
**** right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA<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 parts

Revision as of 14:35, 6 August 2007

Lab schedule

  • mouse insulin gene cloning (delay to 8/7)
    • primer design for [http://www.informatics.jax.org/searches/accession_report.cgi?id=MGI%3A96573 insulin II of Mus musculus], [http://www.ncbi.nlm.nih.gov/entrez/viewer.fcgi?cmd=Retrieve&db=Nucleotide&list_uids=142350788&dopt=GenBank&WebEnv=04iJq7mV5tF1N_LSoJWAy8uc-0i_vXvfkGc3KMxcxnrIPG9K-LDVOuYkZWjzh6BpujGcDGNGQk0k9I%40D4770B146B72B990_0041SID&WebEnvRq=1 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 parts
    • ompR binding ompC promoter
    • [http://partsregistry.org/Part:BBa_C0082 tar-EnvZ fusion construct]: cut EnvZ part for our fusion construct
  • antibiotics LB plate preparation
    • ampicillin
    • kanamycin
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