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Team:TU Delft/21 June 2010 content
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| - | + | =Lab work= | |
| + | |||
| + | ==Characterization of Anderson RBS sequences== | ||
| + | [[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" | ||
| - | | | + | |'''#''' |
| - | |2 | + | |'''Sample''' |
| + | |''' Enzyme 1''' | ||
| + | |'''Enzyme 2''' | ||
| + | |'''Buffer''' | ||
| + | |'''BSA''' | ||
| + | |'''Needed fragment''' | ||
|- | |- | ||
| - | | | + | |1 |
| - | | | + | |1.0 μg pSB1T3 |
| - | + | |EcoRI | |
| - | |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. | |
| - | + | ||
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.
