Brown: Team Resistance/21 October 2008

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(Difference between revisions)
(New page: {{BNBK}} ==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 D...)
(→Restriction Digest of S105 cassette and PVJ4 cassette with EcoRI and SpeI:)
 
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===Restriction Digest of S105 cassette and PVJ4 cassette with EcoRI and SpeI:===
===Restriction Digest of S105 cassette and PVJ4 cassette with EcoRI and SpeI:===
*S105
*S105
-
1 37.5 ul Milli-Q Water  
+
# 37.5 ul Milli-Q Water  
-
2 5 ul Multicore
+
# 5 ul Multicore
-
3 .5 ul BSA
+
# .5 ul BSA
-
4 5 ul DNA
+
# 5 ul DNA
-
5 1 ul EcoRI and 1 ul Spe I
+
# 1 ul EcoRI and 1 ul Spe I
*PVJ4
*PVJ4
-
1 30.5 ul Milli-Q Water  
+
# 30.5 ul Milli-Q Water  
-
2 5 ul Multicore
+
# 5 ul Multicore
-
3 .5 ul BSA
+
# .5 ul BSA
-
4 12 ul DNA
+
# 12 ul DNA
-
5 1 ul EcoRI and 1 ul Spe I
+
# 1 ul EcoRI and 1 ul Spe I
*50 ul total reaction volume
*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:
 +
# PVJ4: 10.5 ng/ul
 +
# S105 Cassette: 9.6 ng/ul
 +
 +
===Ligations===
 +
*6:1 Ligations of S105 Cassette and PVJ4 with Standard Biobrick Vector

Latest revision as of 16:05, 21 October 2008


Toxipop lab notebook.png

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
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