Difference between revisions of "User:Megan E. Garber/Notebook/BioE140L - synthesis of vanillin/Entry Base"

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(Autocreated Lab Notebook name=User:Megan E. Garber/Notebook/BioE140L - synthesis of vanillin/Entry Base, content from MediaWiki:EntryContentDefault)
 
(March 5th 2013)
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==Entry title==
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== March 5th 2013==
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March 5 2013 Synthesis of vanillin part asbF-1
 
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Construction File Mix oligos for one synthon, do PCA1 protocol (517bp, pca1)
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PCR pca1 with asbF-1_oligo_3 and asbF-1_oligo_1 by PCA2 protocol (517bp, pca2)
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Digest pca2 with EcoRI/BamHI (yes, BamHI), gp (472+26+19 bp, pcr_dig)
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(get pre-digested vector from JCA of pBca9145-Bca1144
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Ligate pcr_dig and vect_dig, transform, pick white colonies, map, sequence
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asbF-1_oligo_3 >ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCATCGGTCTCCT
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asbF-1_oligo_1 > AAGTATCTTTCCTGTGCCCAGGATCCATGCCGTCTCATCTAATAAGACGTACA
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Predict the product of each assembly step:
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pca1 ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCATCGGTCTCCTATGAAGTATTCATTGTGTACCATCTCATTTAGGCACCAATTAATCTCATTCACAGATATAGTTCAATTCGCATACGAAAACGGATTTGAGGGTATTGAATTATGGGGAACACATGCTCAGAACTTGTATATGCAAGAGTATGAAACTACAGAAAGGGAATTGAATTGTTTAAAGGATAAGACCTTGGAAATTACTATGA
  
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'''PCA protocol'''[http://openwetware.org/wiki/Template:SBB-PCA]
 
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Revision as of 22:36, 12 March 2013

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March 5th 2013

March 5 2013 Synthesis of vanillin part asbF-1 Construction File Mix oligos for one synthon, do PCA1 protocol (517bp, pca1) PCR pca1 with asbF-1_oligo_3 and asbF-1_oligo_1 by PCA2 protocol (517bp, pca2) Digest pca2 with EcoRI/BamHI (yes, BamHI), gp (472+26+19 bp, pcr_dig) (get pre-digested vector from JCA of pBca9145-Bca1144 Ligate pcr_dig and vect_dig, transform, pick white colonies, map, sequence asbF-1_oligo_3 >ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCATCGGTCTCCT asbF-1_oligo_1 > AAGTATCTTTCCTGTGCCCAGGATCCATGCCGTCTCATCTAATAAGACGTACA Predict the product of each assembly step: pca1 ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCATCGGTCTCCTATGAAGTATTCATTGTGTACCATCTCATTTAGGCACCAATTAATCTCATTCACAGATATAGTTCAATTCGCATACGAAAACGGATTTGAGGGTATTGAATTATGGGGAACACATGCTCAGAACTTGTATATGCAAGAGTATGAAACTACAGAAAGGGAATTGAATTGTTTAAAGGATAAGACCTTGGAAATTACTATGA

PCA protocol[1]