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Team:Newcastle/16 July 2010
From 2010.igem.org
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#*double digests with Pst1 and Xba1 | #*double digests with Pst1 and Xba1 | ||
| - | #Gel electrophoresis - the [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] | + | #Gel electrophoresis - the protocol for [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] was followed. |
Inference | Inference | ||
Revision as of 06:38, 9 August 2010
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LacI BioBrick Construction
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).
- The aim of today's experiment is to screen for the lacI insert. This could be done using PCR but that would require specific primers and is not as reliable.
Protocol
- Miniprep - the miniprep protocol was followed.
- Restriction digests - the restriction digests protocol was followed to allow the length of the DNA to be checked using single and double digest.
- Single digest with Pst1
- double digests with Pst1 and Xba1
- Gel electrophoresis - the protocol for gel electrophoresis was followed.
Inference
- Set up liquid broth culture in LB
   
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