Difference between revisions of "Team:UCLA/Notebook"

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       </div>
 
       </div>
 
       <div class="columns small-9">
 
       <div class="columns small-9">
−
         <h5>Isolation of template DNA</h5>
+
         <h5>PCR of Gibson fragments for EBL1 and EBL2 in pSB1C3</h5>
−
         <p> <b> Miniprep of EC93 and EC869 </b> </p>
+
         <p> 50uL rxn: </p>
 +
        <ol>
 +
          <li>2.5 uL F primer</li>
 +
          <li>2.5 uL R primer</li>
 +
          <li>25 uL Q5 2X MM</li>
 +
          <li>19 uL dH2O</li>
 +
        </ol>
 +
        <p> Fragments: </p>
 
         <ul>  
 
         <ul>  
−
           <li>EC93 #1: 145.42 ng/μL</li>
+
           <li>EBL1 in pSB1C3 frag 2</li>
−
           <li>EC93 #2: 198.85 ng/μL</li>
+
           <li>EBL2 in pSB1C3 frag 2</li>
−
           <li>EC869 #1: 90.81 ng/μL</li>
+
           <li>EBL2 in pSB1C3 frag 3</li>
−
          <li>EC869 #2: 45.95 ng/μL</li>
+
 
         </ul>
 
         </ul>
−
         <p> gDNA isolation of ATCC 13048 </p>  
+
         <p> Cycling conditions: </p>
 
         <ul>
 
         <ul>
−
        <li> 34.87 ng/μL </li>
+
          <li> 98C -- 30s</li>  
 
         </ul>
 
         </ul>
−
         <p> Gibson primers designed for EBL1 and EBL2 toxin switched constructs in pSB1C3 and pSC101</p>
+
         <ul type = "i">
 +
          <p> 28 cycles: </p>
 +
          <li> 98C -- 10s</li>
 +
          <li> 66C -- 30s</li>
 +
          <li> 72C -- 140s</li>
 +
        </ul>
 +
        <ul>
 +
          <li> 72C -- 2min</li>
 +
        </ul>
 +
 
 
       </div>
 
       </div>
 
     </div>
 
     </div>

Revision as of 23:16, 9 August 2016

JOURNAL

Projects

CDI

Protein Cages

Week

1

Construction of ATCC 13048 Loci 2 in pSB1C3

Isolation of template DNA from EC93, EC869, and ATCC 13048; PCR of Gibson fragments for EBL2 in pSB1C3 construct

Jun

27

Inoculation of strains obtained from Dr. Christopher Hayes

Inoculated overnight cultures of:

  • EPI100 w/ EC93 cosmid
  • X90 w/ EC869 cosmid
  • ATCC 13048

Conditions:

  • 10mL LB
  • 10uL ampicillin
  • 37 C for 16h shaking
Jun

28

Isolation of template DNA

Miniprep of EC93 and EC869

  • EC93 #1: 145.42 ng/μL
  • EC93 #2: 198.85 ng/μL
  • EC869 #1: 90.81 ng/μL
  • EC869 #2: 45.95 ng/μL

gDNA isolation of ATCC 13048

  • 34.87 ng/μL

Gibson primers designed for EBL1 and EBL2 toxin switched constructs in pSB1C3 and pSC101

Jun

29

PCR of Gibson fragments for EBL1 and EBL2 in pSB1C3

50uL rxn:

  1. 2.5 uL F primer
  2. 2.5 uL R primer
  3. 25 uL Q5 2X MM
  4. 19 uL dH2O

Fragments:

  • EBL1 in pSB1C3 frag 2
  • EBL2 in pSB1C3 frag 2
  • EBL2 in pSB1C3 frag 3

Cycling conditions:

  • 98C -- 30s

    28 cycles:

  • 98C -- 10s
  • 66C -- 30s
  • 72C -- 140s
  • 72C -- 2min
Week

1

Title For The Week

Description blah blah blah

Jun

24

Title for the day (e.g. "Cloning Mutants")

Today we did cloning. fuck cloning. blah blah blah.