SBB10Ntbk-AmyKristofferson: Difference between revisions
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(New page: <pre> Construction of FokI- basic part PCR ak10F/ak20R Registry part BBa_K243001 (230 bp, gp = A) PCR ak20F/ak30R Registry part BBa_K243001 (71 bp, gp = B)...) |
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Revision as of 23:14, 8 February 2010
Construction of FokI- basic part
PCR ak10F/ak20R Registry part BBa_K243001 (230 bp, gp = A)
PCR ak20F/ak30R Registry part BBa_K243001 (71 bp, gp = B)
PCR ak30F/ak40R Registry part BBa_K243001 (245 bp, gp = C)
PCR ak40F/ak11R Registry part BBa_K243001 (135 bp, gp = D)
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PCR ak10F/ak11R on A+B+C+D (619bp, EcoRI/BamHI)
Digest pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L)
Product is pBjk2741-Bak1234 {<FokI->}
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ak10F Forward EcoRI for FokI- CCATAgaattcCAGagatctCAGCTGGTTaaatctgaactggaggagaa
ak20F Base substitution 1 gtaaaccagACggtgccatt
ak20R Base substitution 1 aatggcaccGTctggtttac
ak30F Base substitution 2 cgttatcgtggAcacaaaagcg
ak30R Base substitution 2 cgcttttgtgTccacgataacgc
ak40F Base substitution 3 caccaatTgcaatggggcag
ak40R Base substitution 3 ctgccccattgcAattggtg
ak11R Reverse BamHI for FokI- CTGATggatccaaaattgatctcgccattg
Construction of ColE2 Basic Part
Digest pBca9145-jtk2642 (EcoRI/BamHI, 2057+485, S)
Sub into pBca9523- Bca1144 (EcoRI/BamHI, 2472+918, L)
Product is pBca9523-jtk2642 {ColE2 ori med}
