To use PCR to extract lacI (promoter, ribosome-binding site (RBS) & coding sequence (CDS)) from plasmid pMutin4 and ligate into vector pSB1AT3 in front of red fluorescent protein (RFP).
Repeat transformation of DH5alpha with ligation mix
Results
The gel image was unfortunately lost, however the gel was run using all of the samples and a PCR product of known length, so as to check that the Molecular weight marker was running correctly. The results were the same as the gel run yesterday, i.e. colonies 1 and 2 contain only pSB1AT3 vector (running at 3 Kbp) and the others contain pSB1AT3 with the RFP insert.
Conclusion
The conclusion that can be reached from these results is that we have not been able to get transformants with the correct plasmid/insert combination. To this end the transformation is repeated. However there is again some confusion concerning the sizes, so the samples we already have will undergo a double digest tomorrow in order to extract whatever inserts may be present.
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