BBa_K572009 1 BBa_K572009 Mouse Beta Carotene Dioxygenase 2011-10-02T11:00:00Z 2015-05-08T01:12:44Z Source Organism : Marine gamma proteobacterium EBAC31A08. We would like to thank Kwang-Hwan "Kevin" Jung, Ph.D., Associate Professor, Department of Life Science and Institute of Biological Interfaces, Sogang University, Korea for sending us the plasmid (pKJ900) with dioxygenase gene. .. false true _743_ 0 7684 9 It's complicated false Forward Primer : 5???TTTACTACGAATTCGCGGCCGCTTCTAGATGGAGATAATATTTG 3??? Reverse Primer : 5???AATCGAACctgcagcggccgctactagtaTTAAAGACTTGAG 3??? Polymerase Chain reactions didn't work for a long time at various temperature conditions. Finally, increasing Magnesium Sulphate in the reaction resulted in bringing about the amplification. The following is the optimized PCR Setup : Template (116 ng/ul) = 0.25 ul 10mM dNTP = 1.0 ul 10x Pfu Buffer (without Magnesium Sulphate) = 2.0 ul + Primer (10 uM) = 1.0 ul - Primer (10 uM) = 1.0 ul Magnesium Sulphate - final concentration of 2.5 - 3 mM Pfu Pol = 1.25 u Nuclease Free Water - make the volume to 20ul false Kousik Sundararajan annotation2151502 1 Start Codon range2151502 1 1 3 annotation2151504 1 Stop Codon range2151504 1 1699 1701 annotation2151503 1 Bc_dioxygenase range2151503 1 1 1701 BBa_K572009_sequence 1 atggagataatatttggccagaataagaaagaacagctggagccagttcaggccaaagtgacaggcagcattccagcatggctgcaggggaccctgctccgaaacgggcccgggatgcacacagtgggagagagcaagtacaaccattggtttgatggcctggcccttctccacagtttctccatcagagatggggaggtcttctacaggagcaaatacctgcagagtgacacctacatcgccaacattgaggccaacagaatcgtggtgtctgagttcggaaccatggcctacccggacccctgcaaaaacatcttttccaaagctttctcctacttgtctcacaccatccccgacttcacagacaactgtctgatcaacatcatgaaatgtggagaagacttctatgcaaccacggagaccaactacatcaggaaaatcgacccccagaccctagagactttggagaaggttgattaccggaagtatgtggcggtaaacctggctacctcgcaccctcattatgacgaggctgggaatgtccttaacatgggcacatccgtcgtggacaaagggaggacaaaatacgtgatatttaagatccctgccacagtgccagacagcaagaagaaagggaagagtcccgtgaagcacgcggaagttttctgctccatttcctcccgctcgctgctctctcccagctactaccacagctttggtgtcacggagaactatgtggtgtttctggagcagccttttaagttggatatcctcaagatggccaccgcatacatgaggggagtgagctgggcttcctgtatgtcattcgacagggaggacaagacatacattcatatcatcgaccagaggaccaggaagcctgtgcctaccaagttctacacagatcccatggtggtcttccatcatgtcaatgcctacgaggaggacggctgtgtgctgtttgatgtgatcgcctatgaggacagcagcctctatcagctcttctacctggccaacctgaacaaggacttcgaggagaagtccaggctgacctcagtgcctaccctcaggaggtttgctgtgcccctccatgtggacaaggatgcagaagtgggctcaaatttagtcaaggtgtcatctacaactgcaacagccctgaaggagaaagacggccatgtctattgccagcccgaggtcctctacgaaggcctagagctccctcggataaattatgcttacaacgggaagccatatcgctacatctttgcagctgaagtacagtggagtccagtcccaaccaagatactgaaatatgacattctcacaaagtcctccttaaagtggtctgaggagagctgctggccagcagagcctctgtttgttcccacgccaggtgcgagggatgaagatgatggagtcattttatcagccatcgtctctacggatccccaaaagctgccttttttactcattctggatgccaaaagttttgcggaactggctcgcgcctctgttgatgcggacatgcacctggaccttcatggtttatttatcccagatgcagactggaatgcagtgaagcagactccagctgaaacgcaagaggttgaaaactcagatcatcccacagatccgacagcaccagaactgagccacagtgaaaatgacttcacagcgggtcatggtggctcaagtctttaa igem2sbol 1 iGEM to SBOL conversion Conversion of the iGEM parts registry to SBOL2.1 Chris J. Myers James Alastair McLaughlin 2017-03-06T15:00:00.000Z