SBB10Ntbk-AmyKristofferson: Difference between revisions
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==Construction Files for my parts== | ==2/17/10: New construction file, set up PCR of A,B,C== | ||
<pre> | |||
Construction of FokI- basic part | |||
PCR gh1000F/gh1001R on BBa_K243001 (261 bp, gp = A) | |||
PCR gh1001F/gh1003R on BBa_K243001 (264 bp, gp = B) | |||
PCR gh1003F/ak11R Registry part BBa_K243001 (141 bp, gp = C) | |||
--------------------------------------------------- | |||
PCR gh1000F/ak11R on A+B+C (619bp, EcoRI/BamHI) | |||
Digest pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) | |||
Product is pBjk2741-sbb22 {<FokI->} | |||
--------------------------------------------------- | |||
gh1000F Forward for part <FokI-! ccagtGAATTCatgAGATCTCAGCTGGTTaaatctgaactggaggag | |||
gh1001F Making internal mutation 1+2 cgttggttccccgatcgattatggcgttatcgtggAcacaaaagc | |||
gh1001R Making internal mutation 1+2 cgccataatcgatcggggaaccaacggtataaatggcaccGTctggtttac | |||
gh1003F Making internal mutation 3 ccatatcaccaatTgcaatggggcagtgctgag | |||
gh1003R Making internal mutation 3 ccattgcAattggtgatatgg | |||
ak11R Reverse BamHI for FokI- CTGATggatccaaaattgatctcgccattg | |||
</pre> | |||
==2/8/10: Construction Files for my parts== | |||
===sbb22=== | ===sbb22=== | ||
<pre> | <pre> |
Revision as of 14:18, 22 February 2010
2/17/10: New construction file, set up PCR of A,B,C
Construction of FokI- basic part PCR gh1000F/gh1001R on BBa_K243001 (261 bp, gp = A) PCR gh1001F/gh1003R on BBa_K243001 (264 bp, gp = B) PCR gh1003F/ak11R Registry part BBa_K243001 (141 bp, gp = C) --------------------------------------------------- PCR gh1000F/ak11R on A+B+C (619bp, EcoRI/BamHI) Digest pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-sbb22 {<FokI->} --------------------------------------------------- gh1000F Forward for part <FokI-! ccagtGAATTCatgAGATCTCAGCTGGTTaaatctgaactggaggag gh1001F Making internal mutation 1+2 cgttggttccccgatcgattatggcgttatcgtggAcacaaaagc gh1001R Making internal mutation 1+2 cgccataatcgatcggggaaccaacggtataaatggcaccGTctggtttac gh1003F Making internal mutation 3 ccatatcaccaatTgcaatggggcagtgctgag gh1003R Making internal mutation 3 ccattgcAattggtgatatgg ak11R Reverse BamHI for FokI- CTGATggatccaaaattgatctcgccattg
2/8/10: Construction Files for my parts
sbb22
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) --------------------------------------------------- PCR ak10F/ak11R on A+B+C+D (619bp, EcoRI/BamHI) Digest pBjk2741-Bca1144 (EcoRI/BamHI, 910+2170, L) Product is pBjk2741-Bak1234 {<FokI->} --------------------------------------------------- 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
sbb36
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}