Difference between revisions of "Part:BBa K1846000:Design"
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To transform ORF-314 sequence into a BioBrick basic part, we had the coding sequence synthesised, including the BioBrick prefix and suffix. The sequence was optimised to remove illegal restriction sites. We restricted both our synthesised ORF-314 and the linearised plasmid backbones (pSB1C3 for shipping, and pSB1K3 for further processing) with EcoRI and PstI restriction enzymes, before ligating the two pieces together using T4 DNA ligase. Success of the cloning procedure was confirmed by restriction with EcoRI and PstI restriction enzymes followed by DNA agarose electrophoresis (Figure 1), with pSB1C3 alone and ORF314 alone as controls. ORF-314 was submitted as BBa_K1846000. | To transform ORF-314 sequence into a BioBrick basic part, we had the coding sequence synthesised, including the BioBrick prefix and suffix. The sequence was optimised to remove illegal restriction sites. We restricted both our synthesised ORF-314 and the linearised plasmid backbones (pSB1C3 for shipping, and pSB1K3 for further processing) with EcoRI and PstI restriction enzymes, before ligating the two pieces together using T4 DNA ligase. Success of the cloning procedure was confirmed by restriction with EcoRI and PstI restriction enzymes followed by DNA agarose electrophoresis (Figure 1), with pSB1C3 alone and ORF314 alone as controls. ORF-314 was submitted as BBa_K1846000. | ||
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Revision as of 20:12, 20 September 2015
Bacteriophage lambda ORF314
- 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
To transform ORF-314 sequence into a BioBrick basic part, we had the coding sequence synthesised, including the BioBrick prefix and suffix. The sequence was optimised to remove illegal restriction sites. We restricted both our synthesised ORF-314 and the linearised plasmid backbones (pSB1C3 for shipping, and pSB1K3 for further processing) with EcoRI and PstI restriction enzymes, before ligating the two pieces together using T4 DNA ligase. Success of the cloning procedure was confirmed by restriction with EcoRI and PstI restriction enzymes followed by DNA agarose electrophoresis (Figure 1), with pSB1C3 alone and ORF314 alone as controls. ORF-314 was submitted as BBa_K1846000.
Source
The gene fragment was synthesised using sequence from the genome of bacteriophage Lambda.