SBB13Ntbk-LarryLi: Difference between revisions
(New page: __NOTOC__ ==~~!~~== ==~~~~== Mix oligos for one synthon, do PCA1 protocol (407 bp, pca1) (Use oligo 1-15 except oligo 12) Use new oligo 1 and oligo 9) Digest pca1 with EcoRI/...) |
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==[[User:Larry Li|Larry Li]] 14:47, 5 March 2013 (EST)== | ==[[User:Larry Li|Larry Li]] 14:47, 5 March 2013 (EST)== | ||
Mix oligos for one synthon, do PCA1 protocol (407 bp, pca1) | Mix oligos for one synthon, do PCA1 protocol (407 bp, pca1) | ||
(Use oligo 1-15 except oligo 12) Use new oligo 1 and oligo 9) | |||
(Use oligo 1-15 except oligo 12) (Use new oligo 1 and oligo 9) | |||
Digest pca1 with EcoRI/BglII, gp (391+11+5 bp, pcr_dig) | Digest pca1 with EcoRI/BglII, gp (391+11+5 bp, pcr_dig) | ||
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Oligo List: | Oligo List: | ||
1 CCOMT-3_oligo_1 TTTGGAATAGAAACAAACATGTCACCTCCGACATGTTGAGACGATGCAGATCTC | 1-CCOMT-3_oligo_1 TTTGGAATAGAAACAAACATGTCACCTCCGACATGTTGAGACGATGCAGATCTC | ||
9 CCOMT-3_oligo_2 TTCCTTACCACCTGGATTGTGGGCTAACATGATTACGTCAATGTGGACTACAC | 9-CCOMT-3_oligo_2 TTCCTTACCACCTGGATTGTGGGCTAACATGATTACGTCAATGTGGACTACAC | ||
10 CCOMT-3_oligo_3 TTAGCCCACAATCCAGGTGGTAAGGAAAGAACACAGAAGGAGTTTGAAGACTT | 10-CCOMT-3_oligo_3 TTAGCCCACAATCCAGGTGGTAAGGAAAGAACACAGAAGGAGTTTGAAGACTT | ||
6 CCOMT-3_oligo_4 GGCTTTGCCTGATAATGGTAAAGTTATTGTTGCCGAGTGCATTTTACCTGTCG | 6-CCOMT-3_oligo_4 GGCTTTGCCTGATAATGGTAAAGTTATTGTTGCCGAGTGCATTTTACCTGTCG | ||
12 CCOMT-3_oligo_5 GGCCAAGGGTGCAGGTTTCCAGGGATTCAAGGTACATTGTAACGCATTTAATA | 12-CCOMT-3_oligo_5 GGCCAAGGGTGCAGGTTTCCAGGGATTCAAGGTACATTGTAACGCATTTAATA | ||
7 CCOMT-3_oligo_6 CTTTAGTTGCTAAAGATGAATCTGGTGCGACAGGTAAAATGCACTCGGCAACA | 7-CCOMT-3_oligo_6 CTTTAGTTGCTAAAGATGAATCTGGTGCGACAGGTAAAATGCACTCGGCAACA | ||
4 CCOMT-3_oligo_7 CTGTCATGATTGGTCTGACGAACATTGCTTGAAATTCTTAAAGAATTGTTATGA | 4-CCOMT-3_oligo_7 CTGTCATGATTGGTCTGACGAACATTGCTTGAAATTCTTAAAGAATTGTTATGA | ||
13 CCOMT-3_oligo_8 CCTTCTTTAAGAACTCCATAATGTATGTATTAAATGCGTTACAATGTACCTTG | 13-CCOMT-3_oligo_8 CCTTCTTTAAGAACTCCATAATGTATGTATTAAATGCGTTACAATGTACCTTG | ||
14 CCOMT-3_oligo_9 CATACATTATGGAGTTCTTAAAGAAGGTTGGATCCTGAGACCTGAGACGGCAT | 14-CCOMT-3_oligo_9 CATACATTATGGAGTTCTTAAAGAAGGTTGGATCCTGAGACCTGAGACGGCAT | ||
8 CCOMT-3_oligo_10 CACCAGATTCATCTTTAGCAACTAAAGGTGTAGTCCACATTGACGTAATCATG | 8-CCOMT-3_oligo_10 CACCAGATTCATCTTTAGCAACTAAAGGTGTAGTCCACATTGACGTAATCATG | ||
3 CCOMT-3_oligo_11 CAATGTTCGTCAGACCAATCATGACAGATCCATTTCATAAAGACGGCATCAGCC | 3-CCOMT-3_oligo_11 CAATGTTCGTCAGACCAATCATGACAGATCCATTTCATAAAGACGGCATCAGCC | ||
0 CCOMT-3_oligo_12 ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCAACATGTCGG | 0-CCOMT-3_oligo_12 ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCAACATGTCGG | ||
5 CCOMT-3_oligo_13 ATAACTTTACCATTATCAGGCAAAGCCTCATAACAATTCTTTAAGAATTTCAAG | 5-CCOMT-3_oligo_13 ATAACTTTACCATTATCAGGCAAAGCCTCATAACAATTCTTTAAGAATTTCAAG | ||
2 CCOMT-3_oligo_14 AGGTGACATGTTTGTTTCTATTCCAAAGGCTGATGCCGTCTTTATGAAATGGAT | 2-CCOMT-3_oligo_14 AGGTGACATGTTTGTTTCTATTCCAAAGGCTGATGCCGTCTTTATGAAATGGAT | ||
11 CCOMT-3_oligo_15 AATCCCTGGAAACCTGCACCCTTGGCCAAGTCTTCAAACTCCTTCTGTGTTCT | 11-CCOMT-3_oligo_15 AATCCCTGGAAACCTGCACCCTTGGCCAAGTCTTCAAACTCCTTCTGTGTTCT | ||
15 CCOMT-3_oligo_16 AAGTATCTTTCCTGTGCCCAGGATCCATGCCGTCTCAGGTCTCAGGATCCAA | 15-CCOMT-3_oligo_16 AAGTATCTTTCCTGTGCCCAGGATCCATGCCGTCTCAGGTCTCAGGATCCAA | ||
PCA Product: | PCA Product: | ||
Revision as of 12:48, 5 March 2013
~~!~~
Larry Li 14:47, 5 March 2013 (EST)
Mix oligos for one synthon, do PCA1 protocol (407 bp, pca1)
(Use oligo 1-15 except oligo 12) (Use new oligo 1 and oligo 9)
Digest pca1 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:
1-CCOMT-3_oligo_1 TTTGGAATAGAAACAAACATGTCACCTCCGACATGTTGAGACGATGCAGATCTC 9-CCOMT-3_oligo_2 TTCCTTACCACCTGGATTGTGGGCTAACATGATTACGTCAATGTGGACTACAC 10-CCOMT-3_oligo_3 TTAGCCCACAATCCAGGTGGTAAGGAAAGAACACAGAAGGAGTTTGAAGACTT 6-CCOMT-3_oligo_4 GGCTTTGCCTGATAATGGTAAAGTTATTGTTGCCGAGTGCATTTTACCTGTCG 12-CCOMT-3_oligo_5 GGCCAAGGGTGCAGGTTTCCAGGGATTCAAGGTACATTGTAACGCATTTAATA 7-CCOMT-3_oligo_6 CTTTAGTTGCTAAAGATGAATCTGGTGCGACAGGTAAAATGCACTCGGCAACA 4-CCOMT-3_oligo_7 CTGTCATGATTGGTCTGACGAACATTGCTTGAAATTCTTAAAGAATTGTTATGA 13-CCOMT-3_oligo_8 CCTTCTTTAAGAACTCCATAATGTATGTATTAAATGCGTTACAATGTACCTTG 14-CCOMT-3_oligo_9 CATACATTATGGAGTTCTTAAAGAAGGTTGGATCCTGAGACCTGAGACGGCAT 8-CCOMT-3_oligo_10 CACCAGATTCATCTTTAGCAACTAAAGGTGTAGTCCACATTGACGTAATCATG 3-CCOMT-3_oligo_11 CAATGTTCGTCAGACCAATCATGACAGATCCATTTCATAAAGACGGCATCAGCC 0-CCOMT-3_oligo_12 ATATAGATGCCGTCCTAGCGAATTCATGAGATCTGCATCGTCTCAACATGTCGG 5-CCOMT-3_oligo_13 ATAACTTTACCATTATCAGGCAAAGCCTCATAACAATTCTTTAAGAATTTCAAG 2-CCOMT-3_oligo_14 AGGTGACATGTTTGTTTCTATTCCAAAGGCTGATGCCGTCTTTATGAAATGGAT 11-CCOMT-3_oligo_15 AATCCCTGGAAACCTGCACCCTTGGCCAAGTCTTCAAACTCCTTCTGTGTTCT 15-CCOMT-3_oligo_16 AAGTATCTTTCCTGTGCCCAGGATCCATGCCGTCTCAGGTCTCAGGATCCAA
PCA Product: CTAGCGAATTCCGTCTCAACATGTCGGAGGTGACATGTTTGTTTCTATTCCAAAGGCTGATGCCGTCTTTATGAAATGGATCTGTCATGATTGGTCTGACGAACATTGCTTGAAATTCTTAAAGAATTGTTATGAGGCTTTGCCTGATAATGGTAAAGTTATTGTTGCCGAGTGCATTTTACCTGTCGCACCAGATTCATCTTTAGCAACTAAAGGTGTAGTCCACATTGACGTAATCATGTTAGCCCACAATCCAGGTGGTAAGGAAAGAACACAGAAGGAGTTTGAAGACTTGGCCAAGGGTGCAGGTTTCCAGGGATTCAAGGTACATTGTAACGCATTTAATACATACATTATGGAGTTCTTAAAGAAGGTTGGATCCTGAGACCTGAGACGAGATCTaaacc
On Thursday we will be doing the PCA.
