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Team:Newcastle/15 July 2010
From 2010.igem.org
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*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. | *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 [[Team:Newcastle/Qiagen_Minipreps|miniprep]] protocol was followed. | #Miniprep - the [[Team:Newcastle/Qiagen_Minipreps|miniprep]] protocol was followed. | ||
#Restriction digests - the [[Team:Newcastle/Restriction_digests| restriction digest]] protocol was followed. | #Restriction digests - the [[Team:Newcastle/Restriction_digests| restriction digest]] protocol was followed. | ||
Revision as of 14:20, 6 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 digest protocol was followed.
- Single digest with Pst1
- double digests with Pst1 and Xba1
- Gel electrophoresis
Set up liquid broth culture in 5 ml of LB - 1 colony from colonies 1-7
   
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