Difference between revisions of "Part:BBa K777000:Design"
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===Design Notes=== | ===Design Notes=== | ||
− | + | Genomic sequence was amplified using the following primers.<br><br> | |
− | Primers: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA. | + | Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA. |
Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´ | Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´ | ||
Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´ | Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´ | ||
+ | <br><br> | ||
+ | <br><br> | ||
+ | One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR. <br><br> | ||
+ | Primers QuikChange: Primers were provided by SIGMA. | ||
+ | |||
+ | Fwd: TarQC for: 5´ ACTGATTGATTACCTAGATTATGGCAATACTGGAG 3´ | ||
+ | |||
+ | Rev: TarQC rev: 5´ TGCCATAATCTAGGTAATCAATCAGTTCAGTTAAC 3´ | ||
===Source=== | ===Source=== |
Revision as of 20:01, 11 September 2012
Tar receptor
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 1282
- 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI.rc site found at 111
Design Notes
Genomic sequence was amplified using the following primers.
Primers PCR: Base pairs in caps represent the respective restriction sites followed by either prefix or suffix. Primers were provided by SIGMA.
Fwd: Tar for + prefix: 5´ actGAATTCgcggccgctTCTAGAtgattaaccgtatccgcgtagtc 3´
Rev: Tar rev + suffix: 5´ tcCTGCAGcggccgctACTAGTcaaaatgtttcccagtttggatc 3´
One XbaI site was found at position 420. The restriction site was changed from TCTAGt to CCTAGt via QuikChange PCR.
Primers QuikChange: Primers were provided by SIGMA.
Fwd: TarQC for: 5´ ACTGATTGATTACCTAGATTATGGCAATACTGGAG 3´
Rev: TarQC rev: 5´ TGCCATAATCTAGGTAATCAATCAGTTCAGTTAAC 3´
Source
The part was amplified from genomic DNA of E. coli str. K-12 substr. DH10B chromosome (NC_010473.1).