User:University of Washington/2 July 2008

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== BioBrick Promoter Measurements ==
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- The four overnight cultures of TOP10 cells containing promoter constructs I20260, I20268, I20269, and I20270 were centrifuged at 14,000 RPM for 3 minutes. Then, the supernatant was decanted out, leaving only pelleted cells.
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- The pelleted cells were miniprepped using a Qiagen miniprep kit.
 +
- 4 uL of each plasmid sequence was added to its own two tubes. One tube had 8 uL of forward primer added to it, and the other had 8 uL of reverse primer added to it. These reaction tubes were submitted to the UW Sequencing Facility for sequencing.
 +
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- The remaining purified plasmid solutions were stored at -20 degrees Celsius.
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 +
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== LuxR from pLac ==
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- I0462 transformed cells failed to grow on Amp plate. I0462 has now failed to transform three times!
 +
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- The I0462 DNA spot from the notebook was nanodropped to verify that it actually contains DNA. It did indeed contain DNA: 55.8 ng/ul, 1.25 (260/280), 1.30 (260/230).
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- Part I763004 was transformed into DH strain, in substition for defective part J04430.
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== LuxR from AraC and TetR ==
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- Nanodrop AraC: 162.5 ng/ul, 1.93 (260/280), 1.47 (260/230)
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== Induction of Conjugative Transfer (Bryan & Scott) ==
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- Designed and ordered primers for Lambda Red recombination of RP4 promoter sequences
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[[Team:University_of_Washington/Notebook#Notebook]]
[[Team:University_of_Washington/Notebook#Notebook]]

Latest revision as of 18:28, 3 July 2008

Contents

BioBrick Promoter Measurements

- The four overnight cultures of TOP10 cells containing promoter constructs I20260, I20268, I20269, and I20270 were centrifuged at 14,000 RPM for 3 minutes. Then, the supernatant was decanted out, leaving only pelleted cells.

- The pelleted cells were miniprepped using a Qiagen miniprep kit.

- 4 uL of each plasmid sequence was added to its own two tubes. One tube had 8 uL of forward primer added to it, and the other had 8 uL of reverse primer added to it. These reaction tubes were submitted to the UW Sequencing Facility for sequencing.

- The remaining purified plasmid solutions were stored at -20 degrees Celsius.


LuxR from pLac

- I0462 transformed cells failed to grow on Amp plate. I0462 has now failed to transform three times!

- The I0462 DNA spot from the notebook was nanodropped to verify that it actually contains DNA. It did indeed contain DNA: 55.8 ng/ul, 1.25 (260/280), 1.30 (260/230).

- Part I763004 was transformed into DH strain, in substition for defective part J04430.


LuxR from AraC and TetR

- Nanodrop AraC: 162.5 ng/ul, 1.93 (260/280), 1.47 (260/230)

Induction of Conjugative Transfer (Bryan & Scott)

- Designed and ordered primers for Lambda Red recombination of RP4 promoter sequences


Team:University_of_Washington/Notebook#Notebook

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