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User:University of Washington/2 July 2008
From 2008.igem.org
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== BioBrick Promoter Measurements == | == BioBrick Promoter Measurements == | ||
| - | - The four overnight cultures of TOP10 cells containing promoter constructs I20260, I20268, I20269, and I20270 were | + | - 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. | - 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. | - 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]] | [[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
