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Team:Newcastle/15 July 2010
From 2010.igem.org
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Screening for lacI insert: | Screening for lacI insert: | ||
| - | # | + | #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. | ||
#*Single digest with Pst1 | #*Single digest with Pst1 | ||
Revision as of 14:14, 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).
Screening for lacI insert:
- 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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