Difference between revisions of "Part:BBa K1336005:Design"
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===Design Notes=== | ===Design Notes=== | ||
When designing PCR primers to change the reverse complement sequence of the ispB gene into the BioBrick format, we truncated the fragment to avoid an illegal PstI restriction site (and so no extra primers were required for site-directed mutagenesis to remove illegal sites). The specific primers were: | When designing PCR primers to change the reverse complement sequence of the ispB gene into the BioBrick format, we truncated the fragment to avoid an illegal PstI restriction site (and so no extra primers were required for site-directed mutagenesis to remove illegal sites). The specific primers were: | ||
| + | <br> | ||
iGM PRE ispB RNAi: 5’ - GTTTCTTC GAATTC GCGGCCGC T TCTAGA G TTA ACG ATC GCG TTG AAC AGC G – 3’ | iGM PRE ispB RNAi: 5’ - GTTTCTTC GAATTC GCGGCCGC T TCTAGA G TTA ACG ATC GCG TTG AAC AGC G – 3’ | ||
| − | iGM SUF REV ispB RNAi:5’–GT TTC TTC CTG CAG CGG CCG CTA CTA GTA ATG AAT TTA GAA AAA ATC AAT GAG–3’ | + | <br>iGM SUF REV ispB RNAi:5’–GT TTC TTC CTG CAG CGG CCG CTA CTA GTA ATG AAT TTA GAA AAA ATC AAT GAG–3’ |
===Source=== | ===Source=== | ||
Latest revision as of 16:15, 21 October 2014
Antisense for octaprenyl diphosphate synthase (ispB gene)
Assembly Compatibility:
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
When designing PCR primers to change the reverse complement sequence of the ispB gene into the BioBrick format, we truncated the fragment to avoid an illegal PstI restriction site (and so no extra primers were required for site-directed mutagenesis to remove illegal sites). The specific primers were:
iGM PRE ispB RNAi: 5’ - GTTTCTTC GAATTC GCGGCCGC T TCTAGA G TTA ACG ATC GCG TTG AAC AGC G – 3’
iGM SUF REV ispB RNAi:5’–GT TTC TTC CTG CAG CGG CCG CTA CTA GTA ATG AAT TTA GAA AAA ATC AAT GAG–3’
Source
The PCR product was obtained from genomic E. coli sequence, via colony boil.