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