IGEM:Cambridge/2008/Notebook/Turing Pattern Formation/2008/09/12: Difference between revisions
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- 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 | - 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 | ||
*Lane2 : HyperladderI | |||
*Lane3 : RBS W1 cut with EcoRI | |||
*Lane4 : RBS W1 cut with XbaI | |||
*Lane5 : RBS W3 cut with EcoRI | |||
*Lane6 : RBS W3 cut with XbaI | |||
*Lane7 : RBS W7 cut with EcoRI | |||
*Lane8 : RBS W7 cut with XbaI | |||
*Lane9 : RBS W9 cut with EcoRI | |||
*Lane10 : RBS W9 cut with XbaI | |||
*Lane11 : RBS W9 uncut | |||
*Lane12 : supercoiled ladder | |||
- Gel2 | |||
*Lane2 : HyperladderI | |||
*Lane3 : RBS S4 cut with EcoRI | |||
*Lane4 : RBS S4 cut with XbaI | |||
*Lane5 : RBS S6 cut with EcoRI | |||
*Lane6 : RBS S6 cut with XbaI | |||
*Lane7 : RBS S8 cut with EcoRI | |||
*Lane8 : RBS S8 cut with XbaI | |||
*Lane9 : RBS S11 cut with EcoRI | |||
*Lane10 : RBS S11 cut with XbaI | |||
*Lane11 : RBS S4 uncut | |||
*Lane12 : supercoiled ladder | |||
==Check PCR products== | ==Check PCR products== |
Revision as of 02:36, 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
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 |