Team:TU Delft/21 June 2010 content

From 2010.igem.org

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<h4>Characterization of Anderson RBS sequences</h4>  
<h4>Characterization of Anderson RBS sequences</h4>  
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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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[[Team:TU_Delft/protocols/restriction_enzyme_digestion|Restriction digestion]] of plasmid backbone pSB1T3.
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
{| style="color:black; background-color:white;" cellpadding="5" cellspacing="0" border="1"
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|'''Digestion reaction'''
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|'''#'''
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|'''Used Buffer'''
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|'''Sample'''
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|''' Enzyme 1'''
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|'''Enzyme 2'''
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|'''Buffer'''
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|'''BSA'''
|'''Needed fragment'''
|'''Needed fragment'''
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|-
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|1,0 μg pSB1T3 + 1 U/μL EcoRI + 1 U/μL PstI
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|1
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|Buffer H (Roche)
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|1.0 μg pSB1T3
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|‘E – ---- – P’
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|EcoRI
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|}
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|PstI
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|H (Roche)
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The digested pSB1T3 was cut from gel and isolate the band.  
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|✗
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|‘E–linear pSB1T3–P’
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|}
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In stead of 10 U/μL restriction enzyme we used 1 U/μL. 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 16:36, 31 July 2010

Lab work

Characterization of Anderson RBS sequences

Restriction digestion of plasmid backbone pSB1T3.

# Sample Enzyme 1 Enzyme 2 Buffer BSA Needed fragment
1 1.0 μg pSB1T3 EcoRI PstI H (Roche) ✗ ‘E–linear pSB1T3–P’

In stead of 10 U/μL restriction enzyme we used 1 U/μL. 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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