Rstclair2012 (Talk | contribs) |
Rstclair2012 (Talk | contribs) |
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<h5>Transformation Procedure</h5> | <h5>Transformation Procedure</h5> | ||
− | + | ◻ added 5uL DNA | |
<br/> | <br/> | ||
− | let sit 30 min on ice | + | ◻ let sit 30 min on ice |
<br/> | <br/> | ||
− | heat | + | ◻ heat shocked at 42C for 30 sec |
<br/> | <br/> | ||
− | + | ◻ placed back on ice for 2 min | |
<br/> | <br/> | ||
− | + | ◻ added 450uL Luria Broth and incubated at 37c for 2 hours | |
<br/> | <br/> | ||
− | + | ◻ plated 100uL on LB/chloramphenicol plates | |
<br/> | <br/> | ||
− | + | ◻ grew overnight at 37C | |
<br/> | <br/> | ||
− | this was done for all 8 machines | + | ◻ this was done for all 8 machines |
<br/> | <br/> | ||
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Picked 2 colonies from each of plate and inoculate it on 5-10 mL LB medium + Chloramphenicol. | Picked 2 colonies from each of plate and inoculate it on 5-10 mL LB medium + Chloramphenicol. | ||
<br/> | <br/> | ||
− | + | Grew the cells overnight (16-18 hours) at 37°C and 220 rpm. | |
<br/> | <br/> | ||
<h6>Day 3:</h6> | <h6>Day 3:</h6> | ||
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◻ Imported data into Excel (Dilution Calculation) Sheet_1 provided | ◻ Imported data into Excel (Dilution Calculation) Sheet_1 provided | ||
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− | ◻ | + | ◻ Diluted the cultures to a target OD600 of 0.02 (see the volume of preloading culture and |
media in Excel (Dilution Calculation) Sheet_1) in 15 ml LB medium + Chloramphenicol | media in Excel (Dilution Calculation) Sheet_1) in 15 ml LB medium + Chloramphenicol | ||
in 50 mL falcon tube (amber, or covered with foil to block light). | in 50 mL falcon tube (amber, or covered with foil to block light). | ||
<br/> | <br/> | ||
− | ◻ | + | ◻ Incubated the cultures at 37°C and 220 rpm. |
<br/> | <br/> | ||
− | ◻ | + | ◻ Took 1 mL samples of the cultures at 0, 2, 4, and 6 hours of incubation. (At each time |
− | point, | + | point, sampled from each of the 8 devices, two colonies per device, for a |
total of 16 samples per time point) | total of 16 samples per time point) | ||
<br/> | <br/> | ||
− | ◻ | + | ◻ Placed samples on ice. |
<br/> | <br/> | ||
− | ◻ | + | ◻ Measured samples (OD and Fl |
− | measurement) | + | measurement) |
<br/> | <br/> | ||
− | ◻ | + | ◻ Recorded data in notebook |
<br/> | <br/> | ||
Revision as of 22:05, 29 September 2017
Florida_Atlantic