Team:TU Delft/21 June 2010 content

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===Lab work===
===Lab work===
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[[Team:TU_Delft/protocols/restriction_enzyme_digestion|Restriction digestion]] of plasmid backbone pSB1T3 (~1,0 μg) with restriction enzymes EcoRI and PstI:
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<h4>Characterization of Anderson RBS sequences</h4>
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{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
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[[Team:TU_Delft/protocols/restriction_enzyme_digestion|Restriction digestion]] of plasmid backbone pSB1T3 using Buffer H of Roche.
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|pSB1T3
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Digestion reaction:                                    needed fragment
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|2,5 μL
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1,0 μg pSB1T3 + 1 U/μL EcoRI and 1 U/μL PstI: ‘E – ---- – P’
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|-
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|Buffer H (Roche)
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|2,0 μL
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|-
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|EcoRI (1 U/μg)
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|1,0 μL
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|PstI (1 U/μg)
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|1,0 μL
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|H2O
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|13,5 μL
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The digested pSB1T3 was cut from gel and isolate the band.  
The digested pSB1T3 was cut from gel and isolate the band.  
However, the gel extraction kit didn’t work as well as we had hoped, it turned out to be the DNA disappearing kit. Tomorrow a new day.
However, the gel extraction kit didn’t work as well as we had hoped, it turned out to be the DNA disappearing kit. Tomorrow a new day.

Revision as of 17:39, 17 July 2010

Lab work

Characterization of Anderson RBS sequences

Restriction digestion of plasmid backbone pSB1T3 using Buffer H of Roche. Digestion reaction: needed fragment 1,0 μg pSB1T3 + 1 U/μL EcoRI and 1 U/μL PstI: ‘E – ---- – P’

The digested pSB1T3 was cut from gel and isolate the band. However, the gel extraction kit didn’t work as well as we had hoped, it turned out to be the DNA disappearing kit. Tomorrow a new day.

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