Difference between revisions of "Team:UCLA/Notebook"
| Line 87: | Line 87: | ||
<h5>PCR of Gibson fragments for EBL1 and EBL2 in pSB1C3</h5> | <h5>PCR of Gibson fragments for EBL1 and EBL2 in pSB1C3</h5> | ||
<p> 50uL rxn: </p> | <p> 50uL rxn: </p> | ||
| − | < | + | <ul> |
<li>2.5 uL F primer</li> | <li>2.5 uL F primer</li> | ||
<li>2.5 uL R primer</li> | <li>2.5 uL R primer</li> | ||
<li>25 uL Q5 2X MM</li> | <li>25 uL Q5 2X MM</li> | ||
<li>19 uL dH2O</li> | <li>19 uL dH2O</li> | ||
| − | </ | + | </ul> |
<p> Fragments: </p> | <p> Fragments: </p> | ||
<ul> | <ul> | ||
| Line 103: | Line 103: | ||
<li> 98C -- 30s</li> | <li> 98C -- 30s</li> | ||
</ul> | </ul> | ||
| − | <ul | + | <ul> <b> |
| − | + | ||
<li> 98C -- 10s</li> | <li> 98C -- 10s</li> | ||
<li> 66C -- 30s</li> | <li> 66C -- 30s</li> | ||
<li> 72C -- 140s</li> | <li> 72C -- 140s</li> | ||
| − | </ul> | + | </b> </ul> |
<ul> | <ul> | ||
<li> 72C -- 2min</li> | <li> 72C -- 2min</li> | ||
</ul> | </ul> | ||
| + | <p> 28 cycles </p> | ||
</div> | </div> | ||
Revision as of 23:17, 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:
- 2.5 uL F primer
- 2.5 uL R primer
- 25 uL Q5 2X MM
- 19 uL dH2O
Fragments:
- EBL1 in pSB1C3 frag 2
- EBL2 in pSB1C3 frag 2
- EBL2 in pSB1C3 frag 3
Cycling conditions:
- 98C -- 30s
- 98C -- 10s
- 66C -- 30s
- 72C -- 140s
- 72C -- 2min
28 cycles
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.
