SBB13Ntbk-LarryLi: Difference between revisions
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Digest pca2 with EcoRI/BglII, gp (391+11+5 bp, pcr_dig) | Digest pca2 with EcoRI/BglII, gp (391+11+5 bp, pcr_dig) | ||
(get pre-digested vector from JCA of pBca9145-Bca1144 (EcoRI/BamHI, vect_dig) | (get pre-digested vector from JCA of pBca9145-Bca1144) (EcoRI/BamHI, vect_dig) | ||
Ligate pcr_dig and vect_dig | Ligate pcr_dig and vect_dig | ||
Revision as of 13:14, 5 March 2013
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Larry Li 14:47, 5 March 2013 (EST)
CCOMT3 Construction File
Mix oligos (1-16) for one synthon, do PCA1 protocol (453 bp, pca1) PCR pca1 with external oligos (Newfoward and Newreverse) by PCA2 protocol (407 bp, pca2) Digest pca2 with EcoRI/BglII, gp (391+11+5 bp, pcr_dig) (get pre-digested vector from JCA of pBca9145-Bca1144) (EcoRI/BamHI, vect_dig) Ligate pcr_dig and vect_dig Oligo List: >CCOMT-3_oligo_1 TTTGGAATAGAAACAAACATGTCACCTCCGACATGTTGAGACGATGCAGATCTC >CCOMT-3_oligo_2 TTCCTTACCACCTGGATTGTGGGCTAACATGATTACGTCAATGTGGACTACAC >CCOMT-3_oligo_3 TTAGCCCACAATCCAGGTGGTAAGGAAAGAACACAGAAGGAGTTTGAAGACTT >CCOMT-3_oligo_4 GGCTTTGCCTGATAATGGTAAAGTTATTGTTGCCGAGTGCATTTTACCTGTCG >CCOMT-3_oligo_5 GGCCAAGGGTGCAGGTTTCCAGGGATTCAAGGTACATTGTAACGCATTTAATA >CCOMT-3_oligo_6 CTTTAGTTGCTAAAGATGAATCTGGTGCGACAGGTAAAATGCACTCGGCAACA >CCOMT-3_oligo_7 CTGTCATGATTGGTCTGACGAACATTGCTTGAAATTCTTAAAGAATTGTTATGA >CCOMT-3_oligo_8 CCTTCTTTAAGAACTCCATAATGTATGTATTAAATGCGTTACAATGTACCTTG >CCOMT-3_oligo_9 CATACATTATGGAGTTCTTAAAGAAGGTTGGATCCTGAGACCTGAGACGGCAT >CCOMT-3_oligo_10 CACCAGATTCATCTTTAGCAACTAAAGGTGTAGTCCACATTGACGTAATCATG >CCOMT-3_oligo_11 CAATGTTCGTCAGACCAATCATGACAGATCCATTTCATAAAGACGGCATCAGCC >CCOMT-3_oligo_12 ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCAACATGTCGG >CCOMT-3_oligo_13 ATAACTTTACCATTATCAGGCAAAGCCTCATAACAATTCTTTAAGAATTTCAAG >CCOMT-3_oligo_14 AGGTGACATGTTTGTTTCTATTCCAAAGGCTGATGCCGTCTTTATGAAATGGAT >CCOMT-3_oligo_15 AATCCCTGGAAACCTGCACCCTTGGCCAAGTCTTCAAACTCCTTCTGTGTTCT >CCOMT-3_oligo_16 AAGTATCTTTCCTGTGCCCAGGATCCATGCCGTCTCAGGTCTCAGGATCCAA >CCOMT3-NewForward ccaaaGAATTCCGTCTCAACATGTCGGAGGTGAC >CCOMT3-NewReverse gactgAGATCTCGTCTCAGGTCTCAGGATCCAAC PCA1 Product: ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCAACATGTCGGAGGTGACATGTTTGTTTCTATTCCAAAGGCTGATGCCGTCTTTATGAAATGGATCTGTCATGATTGGTCTGACGAACATTGCTTGAAATTCTTAAAGAATTGTTATGAGGCTTTGCCTGATAATGGTAAAGTTATTGTTGCCGAGTGCATTTTACCTGTCGCACCAGATTCATCTTTAGCAACTAAAGGTGTAGTCCACATTGACGTAATCATGTTAGCCCACAATCCAGGTGGTAAGGAAAGAACACAGAAGGAGTTTGAAGACTTGGCCAAGGGTGCAGGTTTCCAGGGATTCAAGGTACATTGTAACGCATTTAATACATACATTATGGAGTTCTTAAAGAAGGTTGGATCCTGAGACCTGAGACGGCATGGATCCTGGGCACAGGAAAGATACTT PCA2 Product: ccaaaGAATTCCGTCTCAACATGTCGGAGGTGACATGTTTGTTTCTATTCCAAAGGCTGATGCCGTCTTTATGAAATGGATCTGTCATGATTGGTCTGACGAACATTGCTTGAAATTCTTAAAGAATTGTTATGAGGCTTTGCCTGATAATGGTAAAGTTATTGTTGCCGAGTGCATTTTACCTGTCGCACCAGATTCATCTTTAGCAACTAAAGGTGTAGTCCACATTGACGTAATCATGTTAGCCCACAATCCAGGTGGTAAGGAAAGAACACAGAAGGAGTTTGAAGACTTGGCCAAGGGTGCAGGTTTCCAGGGATTCAAGGTACATTGTAACGCATTTAATACATACATTATGGAGTTCTTAAAGAAGGTTGGATCCTGAGACCTGAGACGAGATCTcagtc
On Thursday we will be doing the PCA.
