Team:Wageningen UR/Notebook/Proj1/June

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(→June - Synchronized Oscillatory System)
(→June - Synchronized Oscillatory System)
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'''June 3'''
'''June 3'''
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Of all incubations, only cultures of BBa_1712019 transformants were made. These were plated on LB medium with Amp.
+
Of all incubations, only cultures of BBa_1712019 transformants have arisen. These transformants were plated on LB medium with Amp.
The yield of miniprepping was determined by using the Nanodrop spectrophotometer. The plasmids were present in the following concentrations:
The yield of miniprepping was determined by using the Nanodrop spectrophotometer. The plasmids were present in the following concentrations:
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'''June 6'''
'''June 6'''
 +
On June 3 was seen BBa_"K325210" and -K325219 transformants did not grow. These have been incubated in the same way as on June 2 again. Also the colony PCR determined transformants with BBa_10462, -E0422, -C0062 and -P0412 in liquid culture (see June 1) were incubated again (BBa_K325210 transformants have not shown growth until this moment). Finally, on June 2 no transformants with BBa_R0062 or -R0063 were found; these have been incubated again too.
-
--1712019--
+
'''June 7'''
 +
 
 +
The liquid cultures were inspected: All incubations resulted in growth. Two transformants had still to be found, which contained BBa_R0040 and -K325909 (see June 2). New colonies were picked out for colony PCRs and to inoculate LB medium in an Eppendorf tube. Afterwards, an electrophoresis gel showed a BBa_R0040 transformant was found.
 +
 
 +
Next to this, transformations with the new BioBrick parts
 +
*BBa_C0261;
 +
*-B0034;
 +
*-C0060;
 +
*-B0015;
 +
*-S0168;
 +
*-C0061 and
 +
*-F2621
 +
were performed.
 +
 +
 
 +
....1712019....

Revision as of 11:44, 31 July 2011

Building a Synchronized Oscillatory System

June - Synchronized Oscillatory System

Jun.png

June 1

12 colonies were screened by colony PCR. A BBa_10462 transformant was used as a positive control and plasmids of BBa_10462 were used in the PCR.

The PCR was confirmed for some of the samples. The corresponding colonies (BBa_10462, -K325210, -E0422, -C0062 and -P0412 transformants) inoculated 10 mL LB medium with Amp and were incubated on a shaker at 37°C overnight.


June 2

With colony PCRs, transformed colonies were found with: BBa_1712019, -"K325210" and -K325219. No transformed colonies were found that should contain: BBa_R0040, -K325909, -R0062 or -R0063. The former colonies were grown in 10 mL LB medium in presence of Amp on a shaker at 37°C overnight.

All inoculations of June 1, but one: the BBa_K325210 transformants, yielded enough growth. The specific colony was incubated, in the same way, again. 5 mL of each of the other cultures was used in miniprepping.


June 3

Of all incubations, only cultures of BBa_1712019 transformants have arisen. These transformants were plated on LB medium with Amp.

The yield of miniprepping was determined by using the Nanodrop spectrophotometer. The plasmids were present in the following concentrations:

  • 95.4 ng/µL of BBa_C0062;
  • 61.0 ng/µL and 70.5 ng/µL of BBa_E0422;
  • 86.2 ng/µL of BBa_I0462 and
  • 86.2 ng/µL of BBa_P0412.


June 6

On June 3 was seen BBa_"K325210" and -K325219 transformants did not grow. These have been incubated in the same way as on June 2 again. Also the colony PCR determined transformants with BBa_10462, -E0422, -C0062 and -P0412 in liquid culture (see June 1) were incubated again (BBa_K325210 transformants have not shown growth until this moment). Finally, on June 2 no transformants with BBa_R0062 or -R0063 were found; these have been incubated again too.


June 7

The liquid cultures were inspected: All incubations resulted in growth. Two transformants had still to be found, which contained BBa_R0040 and -K325909 (see June 2). New colonies were picked out for colony PCRs and to inoculate LB medium in an Eppendorf tube. Afterwards, an electrophoresis gel showed a BBa_R0040 transformant was found.

Next to this, transformations with the new BioBrick parts

  • BBa_C0261;
  • -B0034;
  • -C0060;
  • -B0015;
  • -S0168;
  • -C0061 and
  • -F2621

were performed.


....1712019....


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