NYMU Taipei/Lab Notes/2007 8 6&7
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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] | + | ** 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 | ||
| - | **** left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATG<font color="red"> | + | **** translation (90-110): GIVDQCCTSICSLYQLENYCN |
| - | **** right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA<font color="red">GTTGCAGTAGTTCTCCAGCT</font> | + | **** transcript (341-403): GGCATTGTAGATCAGTGCTGCACCAGCATCTGCTCCCTCTACCAGCTGGAGAACTACTGCAAC, 63 bp |
| + | **** left: <font color="green">GTTTCTTCGAATTCGCGGCCGCTTCTAGATG</font><font color="red">GGCATTGTAGATCAGTGCTG</font> | ||
| + | **** right: <font color="green">GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA</font><font color="red">GTTGCAGTAGTTCTCCAGCT</font> | ||
| + | ****Potential dimer formation! Do NOT use this pair of primers! We need a new design. | ||
*** primer for B chain | *** primer for B chain | ||
| - | **** left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATG<font color="red"> | + | **** translation (25-54): FVKQHLCGSHLVEALYLVCGERGFFYTPMS |
| - | **** right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTA<font color="red">GGACATGGGTGTGTAGAAGA</font> | + | **** transcript (146-235):TTTGTCAAGCAGCACCTTTGTGGTTCCCACCTGGTGGAGGCTCTCTACCTGGTGTGTGGGGAGCGTGGCTTCTTCTACACACCCATGTCC, 90 bp |
| + | **** left: <font color="green">GTTTCTTCGAATTCGCGGCCGCTTCTAGATG</font><font color="red">TTTGTCAAGCAGCACCTTTGT</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 iGEM 2007 parts | ||
| + | ** 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 | ||
| + | * antibiotics LB plate preparation | ||
| + | ** ampicillin | ||
| + | ** kanamycin | ||
| + | [[Image:0806Plates.jpg|300px]] | ||
Latest revision as of 09:18, 17 August 2007
Lab schedule
- 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]
- 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: GTTTCTTCGAATTCGCGGCCGCTTCTAGATGGGCATTGTAGATCAGTGCTG
- right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGTTGCAGTAGTTCTCCAGCT
- Potential dimer formation! Do NOT use this pair of primers! We need a new design.
- primer for B chain
- translation (25-54): FVKQHLCGSHLVEALYLVCGERGFFYTPMS
- transcript (146-235):TTTGTCAAGCAGCACCTTTGTGGTTCCCACCTGGTGGAGGCTCTCTACCTGGTGTGTGGGGAGCGTGGCTTCTTCTACACACCCATGTCC, 90 bp
- left: GTTTCTTCGAATTCGCGGCCGCTTCTAGATGTTTGTCAAGCAGCACCTTTGT
- right: GTTTCTTCCTGCAGCGGCCGCTACTAGTATTATTAGGACATGGGTGTGTAGAAGA
- 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]
- 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
- [http://partsregistry.org/Part:BBa_C0082 tar-EnvZ fusion construct]: cut EnvZ part for our fusion construct
- ompR binding ompC promoter
- antibiotics LB plate preparation
- ampicillin
- kanamycin
