Brown: Team Resistance/21 October 2008

21 October 2008

 * Optical Density S105 gene Restriction Digest with EcoRI and Spe I: 116.4 ng/ul
 * Optical Density S105 cassette after PCR Purification: 108.6 ng/ul
 * Optical Density PVJ4 cassette after PCR Purification: 45.4 ng/ul

Restriction Digest of S105 cassette and PVJ4 cassette with EcoRI and SpeI:

 * S105
 * 1)  37.5 ul Milli-Q Water
 * 2)  5 ul Multicore
 * 3)  .5 ul BSA
 * 4)  5 ul DNA
 * 5)  1 ul EcoRI and 1 ul Spe I


 * PVJ4
 * 1)  30.5 ul Milli-Q Water
 * 2)  5 ul Multicore
 * 3)  .5 ul BSA
 * 4)  12 ul DNA
 * 5)  1 ul EcoRI and 1 ul Spe I


 * 50 ul total reaction volume


 * Multicore has Star Activity and can possibly cut at sites similar to the EcoRI. Should use Buffer E from here on out.  The digest was run for 15 minutes at 37 degrees Celsius.


 * PCR Purification of Restriction Digest performed:
 * Optical Densities:
 * 1) PVJ4: 10.5 ng/ul
 * 2) S105 Cassette: 9.6 ng/ul

Ligations

 * 6:1 Ligations of S105 Cassette and PVJ4 with Standard Biobrick Vector