Revision as of 10:57, 10 August 2010

LacI BioBrick Construction

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).

Restriction digest of Plasmid extracts with

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 ==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).
-*To see whether the LacI insert is in the pSB1AT3 plasmid (same as yesterday).
-==Materials and Protocol==
+==Materials==
-*Performed a gel electrophoresis  with remaining PCR products + Controls with same restriction digests.
+*[[Team:Newcastle/13_July_2010|Plasmid extracts]]
+*Restriction enzymes ''Xba''1 and ''Pst''1
-====Transformation====
-*''E. coli'' DH5α was transformed again with our plasmid. The [https://2010.igem.org/TeamNewcastleTransformation_of_E._coli#Protocol| transformation protocol] was followed.
-====Miniprep====
-*For this step the [[Team:Newcastle/Qiagen Minipreps| Qiagen Minipreps]] protocol was followed. The 7 colonies that grew on the plates from yesterday were cultured ovenight on a shaking shelf at 37°C in universal tubes.
-====Restriction digests====
-*The [https://2010.igem.org/TeamNewcastleRestriction_digests| restriction digest] protocol was followed for this step.
-(we followed Wendy's cards/Phil's lab-book)
+==Protocol==
+Restriction digest of Plasmid extracts with ''
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