SBB13Ntbk-LarryLi: Difference between revisions
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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 ( | Mix oligos for one synthon, do PCA1 protocol (453 bp, pca1) | ||
Digest | PCR pca1 with external oligos 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) | (get pre-digested vector from JCA of pBca9145-Bca1144 (EcoRI/BamHI, vect_dig) | ||
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Oligo List: | 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. | On Thursday we will be doing the PCA. | ||
Revision as of 13:07, 5 March 2013
~~!~~
Larry Li 14:47, 5 March 2013 (EST)
Mix oligos for one synthon, do PCA1 protocol (453 bp, pca1)
PCR pca1 with external oligos 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.
