Revision as of 19:50, 25 October 2010

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-=LacI BioBrick Construction=
+=Research=
-==Aims==
-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).
-==Materials==
-*[[Team:Newcastle/13_July_2010|Overnight digests]]
-*[[Team:Newcastle/8_July_2010|Ligation mix]]
-*Competent ''E. Coli'' DH5α
-==Protocol==
-*The overnight digest are run on a gel
-*[[Team:Newcastle/9_July_2010|Repeat]] transformation of ''E. coli'' DH5α 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 [[Team:Newcastle/13_July_2010|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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Team:Newcastle/14 July 2010