Difference between revisions of "Team:US AFRL CarrollHS/Experiments"

Line 94: Line 94:
 
</ul>
 
</ul>
 
<li>Inoculate 250µL LB into vial</li>
 
<li>Inoculate 250µL LB into vial</li>
 +
<ul>
 +
<li>incubate at 37℃ with shaking for 1.5 hours</li>
 +
</ul>
 +
<li>Plate 1x/9x</li>
 +
</ol>
 +
</p>
 +
 +
<h3>JM109 Cells:</h3>
 +
<p>
 +
<ol>
 +
<li>Thaw vial of JM109 cells on ice</li>
 +
<li>Label 14 mL Falcon tube & plane on ice</li>
 +
<li>Inoculate 50 µL JM109 cells into Falcon tube</li>
 +
<li>Inoculate 2µL DNA into 50µL JM109 cells</li>
 +
<ul>
 +
<li>incubate on ice for 20m</li>
 +
<li>heat shock at 42℃ for 45 seconds</li>
 +
</ul>
 +
<li>Inoculate 950µL LB into vial</li>
 
<ul>
 
<ul>
 
<li>incubate at 37℃ with shaking for 1.5 hours</li>
 
<li>incubate at 37℃ with shaking for 1.5 hours</li>

Revision as of 23:15, 24 October 2017


Experiment

Transformations

BI21 Cells:

  1. Thaw 50 µL vial of BL21 cells on ice
  2. Inoculate 2µL DNA into 50µL BL21 cells
    • incubate on ice for 30 minutes
    • heat shock at 42℃ for 10 seconds
    • incubate on ice for 5m
  3. Inoculate 250µL LB into vial
    • incubate at 37℃ with shaking for 1.5 hours
  4. Plate 1x/9x

JM109 Cells:

  1. Thaw vial of JM109 cells on ice
  2. Label 14 mL Falcon tube & plane on ice
  3. Inoculate 50 µL JM109 cells into Falcon tube
  4. Inoculate 2µL DNA into 50µL JM109 cells
    • incubate on ice for 20m
    • heat shock at 42℃ for 45 seconds
  5. Inoculate 950µL LB into vial
    • incubate at 37℃ with shaking for 1.5 hours
  6. Plate 1x/9x

Methodology