User:Megan E. Garber/Notebook/BioE140L - synthesis of vanillin/Entry Base: Difference between revisions
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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'''[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 2013March 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] | |
