NYMU Taipei/Lab Notes/2007 8 6&7
From 2007.igem.org
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<h3>Lab schedule</h3> | <h3>Lab schedule</h3> | ||
- | * mouse insulin gene cloning ( | + | * mouse insulin gene cloning (8/6 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> | *** sequence for <font color="green">standard assembly (marked in green)</font> and <font color="red">PCR reaction (marked in red)</font> |
Revision as of 13:55, 7 August 2007
Lab schedule
- mouse insulin gene cloning (8/6 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 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
- ompR binding ompC promoter
- antibiotics LB plate preparation
- ampicillin
- kanamycin