IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/09/12: Difference between revisions
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[[Image:gel21092pcr.gif|250px|center]] | [[Image:gel21092pcr.gif|250px|center]] | ||
The size of our plasmid is about 3000bp (ok on the gel) | |||
*RBS W | |||
- W1 with EcoRI : cut | |||
- W1 with XbaI : uncut, self ligation | |||
- W3 with EcoRI : cut | |||
- W3 with XbaI : uncut, self ligation | |||
- W7 with EcoRI : impossible to see | |||
- W7 with XbaI : uncut, self ligation | |||
- W9 with EcoRI : cut | |||
- W9 with XbaI : uncut, self ligation | |||
*RBS S | |||
- S4 with EcoRI : cut | |||
- S4 with XbaI : uncut, self ligation | |||
- S6 with EcoRI : cut | |||
- S6 with XbaI : 2 band, this plasmid is partially cut!!! Transformation with RBS S | |||
- S8 with EcoRI : cut | |||
- S8 with XbaI : uncut, self ligation | |||
- S11 with EcoRI : cut | |||
- S11 with XbaI : uncut, self ligation | |||
==Check PCR products== | ==Check PCR products== |
Revision as of 02:44, 16 September 2008
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RBS screening (single digest)- Plasmid miniprep 6 colonies of RBS S and 6 colonies of RBS W (without endo wash buffer) - Nanodrop
- Single digest : with EcoRI and XbaI, add SDW and DNA (according to the previous table, 300mg of DNA), 2μL of fast digest buffer, and 1μL of enzyme - Gel1
The size of our plasmid is about 3000bp (ok on the gel)
- W1 with EcoRI : cut - W1 with XbaI : uncut, self ligation - W3 with EcoRI : cut - W3 with XbaI : uncut, self ligation - W7 with EcoRI : impossible to see - W7 with XbaI : uncut, self ligation - W9 with EcoRI : cut - W9 with XbaI : uncut, self ligation
- S4 with EcoRI : cut - S4 with XbaI : uncut, self ligation - S6 with EcoRI : cut - S6 with XbaI : 2 band, this plasmid is partially cut!!! Transformation with RBS S - S8 with EcoRI : cut - S8 with XbaI : uncut, self ligation - S11 with EcoRI : cut - S11 with XbaI : uncut, self ligation Check PCR products- Gel (2μL of each product)
- agr B : 2 bands? - agrC : ok - rep : small band but good size - backbones : ok - promoters : ok - RBS : nothing |