Team:TU Delft/21 June 2010 content

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We had some drawbacks today. 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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=Lab work=
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==Characterization of Anderson RBS sequences==
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'''Lab work'''
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[[Team:TU_Delft/protocols/restriction_enzyme_digestion|Restriction digestion]] of plasmid backbone pSB1T3.
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Restriction digestion of plasmid backbone pSB1T3:
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{| 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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|pSB1T3 (~1,0 μg)
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|'''#'''
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|2,5 μL
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|'''Sample'''
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|-
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|''' Enzyme 1'''
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|Buffer H (Roche)
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|'''Enzyme 2'''
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|2,0 μL
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|'''Buffer'''
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|'''BSA'''
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|'''Needed fragment'''
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|-
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|1
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|1.0 μg pSB1T3
|EcoRI
|EcoRI
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|1,0 μL
 
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|-
 
|PstI
|PstI
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|1,0 μL
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|H (Roche)
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|-
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|✗
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|H2O
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|‘E–linear pSB1T3–P’
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|13,5 μL
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|}
|}
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Cut digested pSB1T3 from gel and isolate the band.
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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.
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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.

Latest revision as of 19:18, 2 August 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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