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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 protocol for [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] was followed. | #Gel electrophoresis - the protocol for [[Team:Newcastle/Gel_electrophoresis|gel electrophoresis]] was followed. | ||
| - | + | #Set up liquid broth culture in LB. The protocol for [[Team:Newcastle/Growing_an_overnight_cultures| growing an overnight culture]] was followed. | |
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Inference | Inference | ||
{{Team:Newcastle/footer}} | {{Team:Newcastle/footer}} | ||
Revision as of 06:41, 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.
- Set up liquid broth culture in LB. The protocol for growing an overnight culture was followed.
Inference
   
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