| 8/8/11 |
Digestion: 1 hour and 30 minutes at 37 degrees Celsius
Plating agar stab of ToxR
Dephosphorylation for 1 hour at 37 degrees Celsius
PCR purify
Ligation-
incubated at 16 degrees Celsius overnight.
Summary:
Digestion of positive result sample of previous FGFR biobrick was performed with
EcoRI to verify if true/false positive results.
In parallel, a digestion of Alfredo’s phototaxis receptor and eYFP was performed
using Alfredo’s backbone XbaI and SpeI. This was done to take it
out of the native back bone and eventually ligate into
new vector backbone (pSBIC3). First the part was dephosphorylated and
then ligated into new vector. At previous FGFR and CheZ* (digested parts)
were also ligated into the vector backbone pSBIC3 in
parallel procedure (will incubate overnight). ToxR,
which did not grow previously, was re-inoculated and incubating overnight.
|
| | |
|
8/9/11
|
Transformation of FGFR and CheZ* with pSB1C3 vector, 50 µL DH5a, 2 µL DNA, 37 degrees Celsius overnight, all chloramphenicol
|
| | |
|
8/11/11
|
Incubate overnight in 3 mL LB chlora at 37 degrees Celsius and 220 rpm
|
| | |
|
8/12/11
|
Post- incubation observation:
i2.17.4- through i2.17.10 all show positive result, i2.17.11- did not change
Results:
All incubated colonies grew as expected. I2.17.4 tube seemed less dense than others.
Negative control did not grow as expected.
|
| | |
|
8/13/11
|
Transformation (50 µL DH5a, 2 µL DNA, 37 degrees Celsius overnight, all carb)
|
| | |
|
8/14/11
|
Incubation in 3 mL LB carb overnight at 37 degrees Celsius and 220 rpm |
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